Resin dispensing device

ABSTRACT

A particle dispensing device including a hollow tube and a plunger inserted through the tube is used to transport and dispense solid reagents such as resin. The particle dispensing device can be coupled to a dispenser assembly that is a component in an automated system for processing biological materials.

REFERENCE TO PRIORITY DOCUMENTS

[0001] This claims priority from U.S. Provisional Application Serial No.60/348,745, filed Oct. 26, 2001, and U.S. Provisional Application SerialNo. 60/348,107, also filed Oct. 26, 2001. Both of those applications areincorporated herein by reference in their entirety.

BACKGROUND OF THE INVENTION

[0002] 1. Field Of the Invention

[0003] The invention relates to process line systems and, moreparticularly, to the transfer of materials onto sample plates forlaboratory analysis.

[0004] 2. Description of the Related Art

[0005] Processing of biological materials often involves the automatedtransfer of sample materials onto reaction points for testing andanalysis. Automated processing reduces the amount of time necessary toprocess large numbers of samples. For example, genetic sequencingefforts, such as the Human Genome project, involve processing of largenumbers of samples, and have produced vast amounts of information forbasic genetic research that have lead to advancements in health care anddrug research. With these advances, scientists can move from basicgenomic discoveries to associating specific phenotypes and diseases, andthereby better identify targets for drug development. Genetic sequencinginvolves tests of samples deposited on microarrays, in conjunction with,for example, mass spectrometry testing.

[0006] Microarrays have been used to execute tests on large batches ofgenetic samples to generate phenotype associations and improveinterpretation of the large data sets that result from such tests. Atypical microarray comprises a substrate on which a large number ofreactive points are located. Testing systems typically use a one-inchsquare array, which is often referred to as a chip. Earlier chips haveninety-six reactive points that receive samples for testing, arranged ina grid of eight points by twelve points. More recently, chips have beenproduced with four times that capacity, having a 16×24 grid of 384reactive target locations on the chip substrate. The high capacitymicroarrays permit the screening of large numbers of samples and canreduce reagent costs because each target location is smaller andtherefore requires less reagent to be deposited for testing.

[0007] Samples are usually prepared in a sample material plate, such asa multiple-well tray called a microtiter plate (MTP). A variety ofliquid reagent materials are combined in the wells and are subjected tovarious heating and mixing cycles. The sample preparation typicallybegins with empty MTPs being delivered to a processing station. Thevarious reagents and biological materials are then added. Some of thesample processing may involve heating, cooling, and mixing of theingredients and biological materials while in the wells of the MTP. Manyhigh-throughput systems involve computer controlled robotic arms thatpick up the MTPs, rotate, and place each MTP at the next processingstation. In this way, each MTP is moved along in the sample preparationprocess. Some stations may take more time to complete than others,thereby creating a bottleneck that hinders increased throughput.

[0008] Typically, completed MTPs reach a processing station where thebiological samples are delivered to the chip target locations, usingpins that are dipped into the sample material, which loads the tip ofthe pin. The loaded pin is then touched to a target area on thesubstrate, so that the sample liquid is transferred to the target bycontact deposition. Pin tools can be problematic for high throughputsystems because the pins themselves may have to be changed if differentsample volumes are desired, or if the nature of the liquid sample ischanged.

[0009] High-throughput testing systems typically use an array of pintools to transfer the samples onto the chip target locations. A grid ofpin tools are mounted on a dispensing head, which is lowered over amultiple-well microtiter plate (MTP) at a loading station so all of thepin tools in the array are simultaneously dipped into a respective welland, when the dispensing head is withdrawn, all the pin tools are loadedwith biological samples, or reagents. Thus, with one downward cycle, allthe pin tools are loaded with a sample material. The dispensing head isthen withdrawn from the MTP, and then lowered over a sample chip. Thesample material is then transferred to the target locations on the chipby contact deposition, which is also referred to as printing.

[0010] It should be apparent that, with ninety-six (or even 384) targetlocations in a one-inch square area, alignment of the dispensing headwith the chip is very important to the accurate delivery of samples tothe target locations. Increases in the throughput of biological samplesin an efficient manner requires increasing the number of pins, therebyreducing the number of load-and-print cycles, and also requires veryquick alignment of the dispensing head over the chip, and also requiresrapid movement from the MTP loading station to the chip.

[0011] The dispensing head with an array of pins (i.e., a block of pins)is usually aligned to a predetermined position relative to the locationat which the chips will be delivered for printing. The alignment processis typically a manual process that is performed at the beginning of aprocessing run, such as at the beginning of a work day. Because theblock is in a fixed position relative to the dispensing head, thealignment of the head to the chips should ensure that all of the pinsare aligned to the target locations on a chip. Each time the processingis halted, however, a manual alignment must be performed again to ensureproper alignment and accurate placement of the pins over the chips.

[0012] A processing run may involve thousands of load-and-printdispensing head cycles. It may be necessary to halt a processing run,such as when it becomes desirable to change or replace pins or the pinblock during a processing run, or when the run must be halted for amechanical failure or to check alignment. This causes a disruption inoperation because, to ensure accurate transfer, another manual alignmentmust be performed before proceeding with the processing run.

[0013] The alignment process after a change in pins or a changed pinblock may be especially important because the new pins may be offsetfrom the previously installed pins, relative to the dispensing head.Thus, if no check of alignment with the new pins is performed, the pintips may make contact with the chip at different locations from before,even though the alignment of the dispensing head to the chip has notchanged, or even if the dispensing head alignment has been checked andconfirmed. The samples will not be accurately transferred to the targetlocations on the chip. Thus, changing pins or pin blocks results in notonly a delay because of the alignment process, but also results in amore complicated alignment process, further slowing down the systemthroughput. Although current systems are capable of processing tens ofthousands of samples in a day, even higher throughput systems aredesired. It should be apparent that current alignment techniques cannoteasily support the demands of high-throughput systems.

[0014] The wells on a MTP often contain sample materials that arethemselves the result of several operations, usually involving themixing of solutions and other processing in each of the wells, toprepare the sample materials. Therefore, the wells must have minimumdimensions to physically permit the sample preparation operations tooccur. For a 384-well MTP, the wells are typically spaced apart atapproximately 4.5 mm between well centers. In contrast, the targetlocations on a chip are typically arranged at the minimal spacingdistance that can avoid sample contamination on the chip, typically atapproximately 1.125 mm between target location centers, although otherspacings may be used. Thus, the 384 wells on a MTP must be spacedfarther apart than the 384 wells on a chip.

[0015] In a typical system, the pins of the dispensing head are arrangedin the same spacing as the wells of the MTP, to permit insertion intothe MTP wells and loading of the pin tips. It should be apparent thatnot all of the target locations on a chip can receive their samples atthe same time, given the differential spacing of the pins. Therefore,systems stagger the delivery of sample material with repeated cycles ofloading and printing with the pins in a dispensing head.

[0016] For example, in the spacing described above, the target locationsare at a spacing that is one-fourth the spacing of the pins in a block.Therefore, for a chip having 384 target locations, a dispensing headhaving a 24-pin array of pins in a block must be loaded and printedthrough sixteen cycles of the dispensing head. It would also benecessary to perform a wash and rinse cycle of the pin block, to preventcontamination, between each loading and printing. It often can requireupwards of twelve minutes to complete the loading and printing for a384-target chip. Even a lower capacity 96-target chip would require fourdispensing head cycles, which would require several minutes to complete.

[0017] Therefore, to print on all the target locations with aconventional 24-pin block, the dispensing head must load the pin blockand print onto a first set of twenty-four target locations such thatevery fourth target location along one dimension on the chip is printed(e.g., first, fifth, ninth, and thirteenth column locations). Along theother dimension, the rows, six target locations will be printed,comprising first row, fifth, ninth, and so forth. The pin block mustthen be washed, rinsed, and loaded for the next printing cycle, duringwhich the 24-pin block is positioned over a second group of targetlocations, offset or staggered from the first group, so that the secondgroup may comprise target locations at the second, sixth, tenth, andfourteenth columns, as well as corresponding row locations.

[0018] After the second group is printed, another wash, rinse, and loadcycle is repeated and then the third dispensing head cycle prints thethird, seventh, eleventh, and fifteenth column of target locations, andthen the fourth cycle prints the target locations for the fourth,eighth, twelfth, and sixteenth columns. In this example, the nextdispensing head cycle would print in columns 17, 21, 25, and 17,followed by columns 18, 22, 26, 28, and so forth, repeating thedispensing head cycles until all wells of the 384-well chip are printed.It should be apparent that the current staggered printing operation canbe a bottleneck to increasing the throughput of sample handling systems.

[0019] As noted above, samples are usually prepared in multiple-welltrays called microtiter plates (MTPs). A variety of reagent materialsare combined in the wells and are subjected to various heating andmixing cycles. The sample preparation typically beings with empty MTPsbeing delivered to a processing station. The various reagents andbiological materials are then added. Some of the sample processing mayinvolve heating, cooling, and mixing of the ingredients and biologicalmaterials while in the wells of an MTP. Many high-throughput systemsinvolve computer controlled robotic arms that pick up the MTPs, rotate,and place each MTP at the next processing station. In this way, each MTPis moved along in the sample preparation process. Some stations may takemore time to complete than others, thereby creating a bottleneck thathinders increased throughput.

[0020] Some of the reagent material may comprise a suspension of liquidand particles mixed together. It is important for the suspensions tohave good mixing of liquid and particles, or solid matter, to ensureproper reactions in the MTP wells. This requirement can make workingwith suspension for MTP wells difficult to work with, because it may bedifficult to keep the suspension adequately mixed and agitated withoutdamaging the particles from excessive mixing and agitation. That is,suspension mixtures can be very unstable and it can be difficult tomaintain them in a sufficiently suspended state.

[0021] An alternative to using a suspension mixture is to keep theparticles separate from the liquid until the suspension mixture isneeded. When it is necessary to mix the particles (which are typicallyin the form of a powder), the particles are deposited into wells of adry particle tray, where each particle well has a predetermined volumeaccording to the laboratory process being performed. Any excess particlematerial that is mounded over the top of any particle well is scrapedoff the top surface of the tray and into a particle reservoir. Theparticle tray is then quickly inverted over the microtiter plate so thatthe contents of each particle well fall into a corresponding well of themicrotiter plate. The particle tray can be tamped with a solid object todislodge any remaining portions of particle matter, ensuring that theproper volume of particle matter is delivered, and then the liquid andparticle contents in each MTP well can be mixed to form the requiredsuspension.

[0022] Maintaining ingredients in powder form can be advantageous,because the solid particles have greater stability and shelf life than acorresponding suspension would have, and keeping the materials in thesolid state avoids the problem of keeping the suspension agitated, butthe particle mixing operation described can be an excessively manualprocess. There is a continuing need for high-throughput biologicalprocessing systems. Such systems are becoming increasingly automated,with processing for tens of thousands of samples each working day. Themanual processing associated with keeping solid particle material out ofsuspension until needed becomes a bottleneck to increased throughput. Itshould be apparent that there is a need for improved techniques forproviding the suspension in MTP wells at the required time duringprocessing of sample materials, to provide greater stability ofmaterial, reduce concerns regarding handling of suspension, and improvecompatibility with increased automation systems.

[0023] Another stumbling block to increasing throughput is therequirement for some systems to perform temperature bath, referred to asthermal cycling. In a typical thermal cycling operation, an MTP plate isplaced on top of a metal plate that conforms to the underside of theMTP. The temperature of the metal plate is controlled through coolingand heating cycles, as desired, thereby affecting the contents of theMTP wells. For high-throughput systems, it is important to ensuregreater heat transfer rates for faster sample processing. It is alsoimportant to achieve greater uniformity of temperature cycling to ensurehighly reproducible biological reactions giving clinically validatedresults.

[0024] Thus, there is a need for improved techniques for alignment ofpins to target locations, for printing between MTP wells of one spacingto target locations at a different spacing that support higherthroughput rates, for particle dispensing, and for thermal cyclingoperations to support increased throughput rates. The present inventionfulfills this need.

SUMMARY

[0025] The present invention is directed toward a particle dispensingdevice that can be used to collect, transport, and dispense solidmaterial, such as reagents, particularly biological and chemicalreagents, within an automated sample handling process line. The particledispensing device is a hollow tube having a proximal end with anopening, a distal end with an opening, and an inside surface. The hollowtube can be any shape suited for use in a process line, such as a rightcircular cylinder, a cone, a frustocone, a cube, a prism, a pyramid, orany other shape that is suitable for coupling to a particle dispensingassembly of a process line. A plunger is slideably and coaxiallyinserted through the proximal end of the hollow tube. The plunger can behollow or solid, and is shaped to fit coaxially within the hollow tube.The proximal end of the particle dispensing device can be designed forcoupling to an array of particle dispensing devices, such as thosedescribed herein.

[0026] In one embodiment, the proximal end of the particle dispensingdevice is coupled to a flat ejection plate that is sized to accommodatemore than one particle dispensing device, preferably up to 384 particledispensing devices. The particle dispensing devices can be arranged in agrid of sixteen devices by twenty-four devices, or in any other mannerthat is suitable for a high-throughput automated processing system. Morepreferably, the flat plate can be sized to accommodate up to 1536particle dispensing devices, which can be arranged in a grid of 32devices by 48 devices, or any other manner that is suitable for ahigh-throughput automated processing system.

[0027] The hollow tube and the plunger can be coaxially integrated in amanner such that a space is formed between the distal end of the plungerand the distal end of the hollow tube. The space can define a volumethat is predetermined and set prior to using the device. The volume canbe increased by moving the plunger away from the distal end of thehollow tube, and it can be decreased by advancing the plunger toward thedistal end of the hollow tube. The volume can be set based on the amountof reagent required by the protocol.

[0028] The methods of the present invention relate to the collection,transport and deposit of reagents, particularly biological reagents,such as resin. These methods are particularly useful in high-throughputautomated processing systems that can be used to perform thousands, oreven millions, of biological reactions per day.

[0029] In one particularly innovative aspect of the invention, aparticle dispensing device, such as the one described above, isprovided. The plunger is set coaxially within the hollow tube so that apredetermined volume of space is formed between the distal end of theplunger and the distal end of the hollow tube. The particle dispensingdevice is then positioned over a reservoir containing a quantity ofsolid reagent, such as resin, or any other biological or chemicalreagent. The hollow tube is then forced into the reservoir, thus forcingparticles of the reagent into the space at the distal end of the hollowtube. The particle dispensing device is then removed from the reservoir,and the particles of reagent remain in the space and are frictionallyengaged with the inside surface of the hollow tube. The particledispensing device is then positioned over a microtiter plate, and theplunger is used to displace the space occupied by the particles ofreagent, thus forcing the reagent out of the space and into themicrotiter plate.

[0030] Other features and advantages of the present invention should beapparent from the following description of the preferred embodiment,which illustrates, by way of example, the principles of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

[0031]FIG. 1 shows a process line system constructed in accordance withthe present invention.

[0032]FIG. 2 is a top view of a microtiter plate that is moved along theprocess line system illustrated in FIG. 1.

[0033]FIG. 3 shows a schematic top view of an exemplary module of theFIG. 1 process line system.

[0034]FIG. 4 is a detail top view of a chip, comprising a substrate withreaction target deposits that will receive sample material from themicrotiter plate illustrated in FIG. 2.

[0035]FIG. 5 is a top view of a multiple-chip holder, containing tenchips of the type illustrated in FIG. 4.

[0036]FIG. 6 is a perspective view of a treatment station of the FIG. 1process line at which sample transfer from microtiter plates to chipstakes place.

[0037]FIG. 7 is a perspective view of a pin array of the treatmentstation shown in FIG. 6.

[0038]FIG. 8 is a view looking down through the reference plateillustrated in FIG. 6, showing the lens of the upward-looking camera.

[0039]FIG. 9A is a view looking up through the reference plate andobserving the underside of the dispensing head illustrated in FIG. 6.

[0040]FIG. 9B is a view from the perspective of the downward-lookingcamera illustrated in FIG. 6, looking down at a chip that is positionedbelow the camera and the dispensing head.

[0041]FIG. 10 is a flow diagram that shows the alignment process for thesystem illustrated in FIG. 1.

[0042]FIG. 11 is a side view of a pin block illustrated in FIG. 5,showing the pins at fully extended pitch.

[0043]FIG. 12 is a top view of the pin configuration illustrated in FIG.11.

[0044]FIG. 13 is a side view of the pin block illustrated in FIG. 11,showing the pins at their fully reduced pitch.

[0045]FIG. 14 is a top view of the pin configuration illustrated in FIG.11.

[0046]FIG. 15 is a perspective view of a portion of the pins illustratedin FIG. 12.

[0047]FIG. 16 is a sequence of schematic representations showing the pinblock of FIG. 11 as it is changed from the fully extended pitch to thefully reduced pitch.

[0048]FIG. 17A is a perspective view of a resin dispensing module of theFIG. 1 processing line showing the dispensing operation and thecompactor.

[0049]FIG. 17B is a perspective view of the resin dispensing module ofFIG. 17A from a different angle.

[0050]FIG. 17C is a top view of the resin dispensing module of FIG. 17A.

[0051]FIG. 17D is a view of the resin dispensing module from a differentperspective from that of FIG. 17A.

[0052]FIG. 18A is a perspective view of the resin dispensing assembly ofthe resin dispensing module of FIG. 17A.

[0053]FIG. 18B is a side elevation view of the resin dispensing assemblydepicted in FIG. 18A.

[0054]FIG. 19A is an exploded three-dimensional view of the resinreservoir of the resin dispensing module of FIG. 17A.

[0055]FIG. 19B is a perspective view of the resin reservoir assembly ofthe resin dispensing module of FIG. 17A.

[0056]FIG. 20A is a schematic representation of the lower portion of onehollow tube in the 384-tube array illustrated in FIG. 17A, showing thetube plunger in its raised position.

[0057]FIG. 20B, is a schematic representation of the FIG. 20Aillustration, with the plunger in its lowest position.

[0058]FIG. 20C shows the hollow tube of FIGS. 20A and 20B carryingparticles of resin, and coupled to a flat ejection plate.

[0059]FIG. 21 is a representation of a computer such as can be used toperform the control tasks described herein.

[0060]FIG. 22 shows a schematic, top view diagram of a process linethermal cycling module where thermal cycling of one or more microtiterplates can be performed.

[0061]FIG. 23 is a schematic side view of a thermal cycling system ofthe thermal cycling module, showing various components of the thermalcycling system.

[0062]FIG. 24 is a schematic side view of a microtiter plate assembly,showing a fluid flow path through which fluid can flow through themicrotiter plate assembly during thermal cycling.

[0063]FIG. 25 shows a perspective view of an exemplary microtiter plate.

[0064]FIG. 26 shows a cross-sectional view of the microtiter plate ofFIG. 25 along the line 25-25 of FIG. 25.

[0065]FIG. 27 shows a perspective view of an exploded flow cell assemblyof a microtiter plate assembly.

[0066]FIG. 28 shows a perspective view of the assembled flow cellassembly.

[0067]FIG. 29 shows a top view of the assembled flow cell assembly.

[0068]FIG. 30 shows a cross-sectional view of the flow cell assemblyalong line 29-29 of FIG. 28.

[0069]FIG. 31 is a cross-sectional view of the flow cell assembly alongthe line 30-30 of FIG. 28.

[0070]FIG. 32 is a bottom view of the flow cell assembly.

[0071]FIG. 33, shows a cross-sectional view of the microtiter plateassembly, showing the microtiter plate positioned in the upper cavity ofthe flow cell assembly.

[0072]FIG. 34 shows a cross-sectional view of the microtiter plateassembly, showing the microtiter plate positioned in the upper cavity ofthe flow cell assembly, the view being along the length of one of theflow channels.

[0073]FIG. 35 is a cross-sectional view of the microtiter plateassembly, looking downward along the line 34-34 of FIG. 30 and showingan inlet cavity and an outlet cavity.

[0074]FIG. 36 shows a cross-sectional view of the microtiter plateassembly coupled to an inlet pipe and an outlet pipe and shows the flowof fluid into the microtiter plate assembly.

[0075]FIG. 37 which shows a downward-looking view of the inlet cavityand the outlet cavity of the microtiter plate assembly, and shows thefluid flow path through the cavities.

[0076]FIG. 38 shows a cross-sectional view of the microtiter plateassembly and shows a fluid flow path.

[0077]FIG. 39 shows a top view of the microtiter plate assembly andshows a fluid flow path.

DETAILED DESCRIPTION

[0078] Unless defined otherwise, all technical and scientific terms usedherein have the same meaning as is commonly understood by one of skillin the art to which this invention belongs. All patents, patentapplications, published applications and publications, Genbanksequences, Websites, and other published material referred to throughoutthe entire disclosure herein are, unless noted otherwise, incorporatedby reference in their entirety.

[0079]FIG. 1 shows a computer controlled process line 100 that isconstructed in accordance with the present invention. The line issometimes referred to as an Automated Processing System 100, and iscontrolled by a computer system 101 that keeps track of microtiterplates (MTPs) as they move along the process line. The computer system101 also controls processing of the MTPs as the MTPs move throughvarious process line modules and work stations. The computer system 101can be used to specify particular modules that the MTPs will be directedto as the process line 100 transports the MTPs. The process line 100includes a plurality of modules or workstations 112, 114, 116, 118, 120,122, and 124 that are connected by a conveyor line 110. As describedbelow, the conveyor line 110 can be used to transport an MTP to all orsome of the modules, where various procedures or processes can beperformed on biological samples of the MTP.

[0080] A module or station whose processing follows that of a priorprocess will be referred to as being “downstream” of the prior process.As will be described further below, the control system of the processline permits a modular configuration that enables extension of theprocess line by inserting new modules before, after, or in between anyof the modules described herein, and also enables extension of theprocess line by adding more stations at any one of the modules, so thata module that performs a specified processing task may have a greater orlesser number of stations that perform that same task, changing innumber as the processing needs require. Thus, it should be appreciatedthat the process line 100 shown in FIG. 1 is merely exemplary withrespect to the quantity of modules, and that the process line 100 couldinclude additional modules or less modules. Furthermore, the processline 100 can include modules where processes other than those describedherein can be performed.

[0081] In the exemplary embodiment shown in FIG. 1, the modules of theprocess line 100 include an introduction module 102, where an MTP can beloaded onto the process line. The introduction module 102 can be used toperform various set-up procedures on the MTP in order to prepare the MTPfor processing in the other modules. The introduction module 102, aswell as other exemplary modules of the process line 100, are describedin more detail below. The introduction module 102 is connected to a lift104 that can upwardly transport the MTP to a bridge 106 that leads tothe remainder of the process line 100. The bridge connects to a secondlift 107 that downwardly transports the MTP to the conveyor line 110,which can transport the MTP to the other modules of the process line100. The conveyor line 110, bridge 106, and lifts 104, 107 include atransport mechanism, such as a conveyor belt, that can support an MTPand move the MTP to each of the modules of the process line 100.

[0082] The lift 104 and bridge 106 permit independent movement ofpersonnel around the MTP introduction module 102 and the processingstations that are downstream of the bridge 106. This permits differentpersonnel to access the first station 102 as compared with the rest ofthe process line 100. In addition, the bridge 106 spatially separatesthe introduction module 102 from the remainder of the process line 100,permitting the use of different materials and maintenance for the twodifferent sections of the process line. Thus, the module 102 can beenvironmentally isolated from the rest of the process line 100, asdescribed in more detail below, in order to overcome any potential riskof sample cross-contamination.

[0083] The processing line can move MTPs along the modules so that MTPprocessing is not entirely sequential or simply batch processing. Thatis, MTPs are received at the first module 102 for processing and arethen moved from module to module, but an MTP can be moved from onemodule to the next as soon as the MTP has completed its processing, sothat an MTP does not necessarily move from one module to the next in theexact same sequence that the MTPs were received at the introductorymodule 102. Thus, modules that take a greater amount of time to processa single MTP may be provided with multiple work stations, such thatmultiple MTPs may be processed at that module. It should be understoodthat any one of the modules 112, 114, 116, 118, 120, 122, 124 mayinclude multiple work stations. That is, each module performs aspecified operation or task associated with biological or chemicalprocessing of sample materials, and each module may include one or morework stations, each of which performs the operations or tasks associatedwith the module. An MTP can bypass a module completely if no processingat that module is needed for that MTP. This increases throughput andincreases the efficiency of the process line 100.

[0084]FIG. 2 shows a top, plan view of a microtiter plate 202 such ascan be processed by the process line. FIG. 2 shows the MTP 202 as a highcapacity plate that contains three hundred eighty-four wells 204,arranged in a grid of sixteen wells by twenty-four wells. Those skilledin the art will understand that MTPs with other capacities are alsoavailable, such as the commonly used ninety-six well MTP, which haswells arranged in eight rows of twelve wells each.

[0085] Process Overview

[0086] The process line 100 comprises a fully integrated continuousbiological processing operation that utilizes combinations of microtiterplates and microtiter plate-sized chip holders to process and transportbiological samples and materials. The process line 100 utilizes athermal-cycling device and procedure, described below, which reduceprocessing time over conventional thermal cyclers. The process line alsoutilizes a nanoliter dispensing device having a dispensing system thatcan be used with microtiter plates and chips of different sizes. Inaddition, the process line uses a resin dispensing device and methodthat permits the addition of dry particulates to an MTP in a rapidmanner. The aforementioned devices are described below in more detail.

[0087] As discussed above, biological reactions are conducted in plasticmicrotiter plates (MTP). The standard commercially available MTPs haveare of 96-well or 384-well configuration, while it is anticipated thatfuture versions will be of 1536-well configuration. The process line 100is configured to accept MTPs of any format. For example, the processline 100 can process MTPs that conform to the Mass EXTEND(hME™)protocol, which has been developed by Sequenom, Inc. of San Diego,Calif. Such MTPs are referred to herein as EXTEND Cocktail plates. Amicrotiter plate is set-up at the beginning of the process line by arobotic arm and microfluidic dispensing equipment, which are located atthe module 102.

[0088] An MTP, such as an MTP containing DNA samples, is set up at theintroduction module 102 and is used to amplify specific target regionsof genomic or plasmid DNA contained in the wells of the MTP. The sameMTP can be used for all subsequent reactions in the process line 100. Ata final module of the process line 100, the products of these reactionsare transferred to one or more microarray chips suitable for conductingmass spectroscopic analysis. The MTPs are initially prepared bydepositing combinations of DNA samples, region-specific amplificationoligonucleotides, and appropriate amplification enzymes and buffers intowells of the MTPs. The MTPs are preferably identified with a magnetic oroptical bar code symbol, sealed and passed into an amplification moduleof the process line, where a process such as PCR is performed. Theintroduction module 102 can also be used to prepare “EXTEND Cocktail”plates, which contain all appropriate reagents, nucleotidetri-phosphates, enzymes and oligonucleotides necessary to conduct theprescribed genotyping analysis.

[0089] The computer system 101 includes tracking software that can beused to define and keep track of the nature of all MTPs introduced intothe introduction module 102. The tracking software can also be used tospecify the process line modules that the MTPs must be transferred toand how the contents of the MTPs will be subsequently used or processed.The bar code of each plate is tracked throughout the progress of theplate through the process line.

[0090] A process line operator can operate the computer 101 thatcontrols operation of the process line 100. The computer 101 can receivefrom the operator operating parameters, commands, and other input thatwill determine the processing of MTPs contained in the process line. Ingeneral, preparing the line 100 for operation involves some preliminaryanalysis to obtain the optimal operating configuration. The following isan overview of the information and data flow used in controllingoperation through the computer 101.

[0091] An operator begins by entering experimental design parametersthrough a software interface program executing in the computer 101. Inone embodiment, the software interface comprises a LaboratoryInformation Management System (LIMS) which is a software interfaceprogram manufactured by Sequenom, Inc. of San Diego, Calif., todetermine the assays that will go on which sample plates. The softwarecan keep track of the contents of MTPs using bar codes that areassociated with each MTP. The operator can initially coordinate the barcode of an MTP to the contents and processes of the MTP using thecomputer 101. For example, barcodes of plates, primers, reagents, hotelplate/reagent holder locations, and module stops, can be read into thesoftware during set-up, such as using a conventional bar code readerthat is coupled to the computer system 101. The software can also obtaindata from the modules of the process line 100 as the MTPs aretransported through the process line. The software is configured tocreate a daily task list for the operator.

[0092] The software creates a work list file for the set-up platform102. The work list can contain, for example MTP set-up information, suchas data regarding the barcode for an MTP and information regarding themodules that the MTP will visit while on the conveyor line 110. Thecomputer system accepts user inputs that define which modules aparticular MTP will be transported to on the process line 100, as wellas which modules will be bypassed. Based on the user inputs, thecomputer system adjusts the movement of the MTP along the process lineso that the MTP is transported to only those modules that are to handlethe biological sample contained in the MTP, and so that the MTP bypassesany module that should not handle the biological sample.

[0093] The Process Line

[0094] The process line 100 is configured to conduct a plurality ofbiological reactions. In one embodiment, the process line 100 conductsover 100,000 individual biological reactions per day and is readilyscaleable to 1,000,000 reactions per day. In another embodiment, theprocess line 100 conducts over 200,000 individual biological reactionsper day. Thus, when used in conjunction is with MTPs having a 384-wellconfiguration, the process line 100 can process up to 520 MTPs per daywhere there are 200,000 individual biological reactions per day and upto 140 MTPs per day where there are 200,000 individual biologicalreactions per day. The configuration is sometimes described herein inthe context of implementing analysis of Single Nucleotide Polymorphisms(SNPs) using the homogeneous Mass EXTEND(hME™) protocol, which has beendeveloped by Sequenom, Inc. of San Diego, Calif. Other configurationsusing the same unit operations but in different combinations arepossible and these will enable other nucleic acid based analyses.

[0095] As mentioned, the process line 100 includes a plurality ofmodules where one or more processes can be performed on an MTP that hasbeen loaded onto the process line 100. An exemplary module 112 is nowdescribed with reference to FIG. 3, which shows a schematic top view ofthe generic module 112. The module 112 includes one or more moduleconveyor lines 301, which are situated transverse to the main conveyorline 110. Each module conveyor line 301 accepts an MTP from the mainconveyor line 110 and transports the MTP to one or more workstations 302that are situated along the module conveyor line 301. For example, theMTP can be transported along the main conveyor line 110 in a directionrepresented by the arrow labeled 303. The MTP can then be moved to themodule conveyor line 301 at the location where the main conveyor line110 meets the module conveyor line 301. The module conveyor line 301 cantransport the MTP to a workstation 302 along a direction represented bythe arrow 305. As described below, the workstation 302 can comprise adevice that performs an automated process on the MTP or on the samplesthat are contained in the wells of the MTP. FIG. 3 shows a singleworkstation 302 situated along each module conveyor line 301, althoughit should be appreciated that the module 112 can include any number ofworkstations 302.

[0096] Thus, as mentioned above, it should be understood that any one ofthe modules 112, 114, 116, 118,120, 122, 124 may include multiple workstations. That is, each module performs a specified operation or taskassociated with biological or chemical processing of sample materials,and each module may include one or more stations, each of which performsthe operations or tasks associated with the module. For example, asshown in FIG. 1, the last module 124 includes stations designated as 124a, 124 b, 124 c to indicate, for example, that multiple water addition,resin mixing, and chip printing stations are provided.

[0097] As mentioned, the MTPs are fitted with one or more barcodes thatcan be utilized to identify the MTP, such as to identify the contents ofthe MTP or the procedures to be performed on the MTP. The barcodes canalso be used to sort data that is associated with each MTP. Thus, themodule 112 can have a conventional barcode reader 308 that is located atthe entrance to each module, as schematically shown in FIG. 3. Inaddition, each module can include a weight measuring device, such as abalance 310, that can be used to measure the weight of each MTP thatenters the module. The balance 310 can be used to measure the weight ofthe MTP before and after processes have been performed on the MTPs inorder to identify a difference in weight of the MTP. A difference inweight could indicate, for example, whether excessive evaporation hasoccurred during thermal processes or whether required reagents have notbeen added to the MTP. The weight measurement at a particular module canalso be used as a reference for future measurements and calculations.

[0098] Exemplary Modules

[0099] An overview of several exemplary modules that can be used in theprocess line 100 is now provided. As shown in FIG. 1, the introductionmodule 102 is situated at the beginning of the process line 100. Theintroduction module 102 is used to initially insert an MTP onto theprocess line 100. In this regard, the process line 100 can include atransport, such as a conveyor belt or a track, that runs the length ofthe process line, such as along the length of the lift 104, bridge 106,lift 107, and conveyor line 110. The MTP is placed on the conveyor atthe introduction module 102. The introduction module 102 can include adevice that seals the wells of the MTP, such as by using an aluminumfilm. Once the MTP has been placed on the conveyor belt, an operator canuse a user interface on the computer system 101 to notify the processline 100 that the MTP is ready for processing.

[0100] The introduction module 102 can be used to prepare and distributematerials to the MTP. For example, the sample material can be a cocktailthat has been or will be subject to a reaction process, such as thePolymerase Chain Reaction (PCR) or to some other reaction process, suchas the “MassEXTEND” reaction process, which is a DNA Polymeraseextension reaction where the oligonucleotide primer is extended throughthe diagnostic region of interest by several bases. A particular MTP canbe selected for use with the introduction module 102.

[0101] With reference to FIG. 1, the lift 104 transports the MTP fromthe module 102 in an upward direction to the bridge 106. The bridge thentransports the MTP to the second lift 107, which then lowers the MTP tothe conveyor line 110. In one embodiment, the introduction module 102 iscontained within a clean room 119 (represented by a dashed box inFIG. 1) that separates the introduction module 102 from the rest of theprocess line 100. The clean room 119 can be sealed, for example, with anairlock to prevent contamination from entering the clean room 119.

[0102] After the lift 104, bridge 106, and lift 107 have transported theMTP from the introduction module 102, the conveyor line 110 receives theMTP from the lift 107. The conveyor line 110 then successivelytransports the MTP to one or more of the modules along the process line100. In an exemplary embodiment, the modules are arranged in the orderdescribed herein, although it should be understood that modules may beadded and deleted while still permitting efficient operation undercontrol of the computer system 101.

[0103] The module 112 is not used for any particular processes in thedescribed embodiment. Rather, module 112 serves as a “virtual” module inthat the module can be used for future expansion. This illustrates theadvantageous modularity of the process line 100, in that modules can beadded, deleted, left empty, expanded or reduced, without affecting theoperation of other modules in the line.

[0104] Module 114 follows module 112 along the process line 110. Module114 comprises an amplification module that includes a thermal cyclingwork station that can be used to thermal cycle the contents of the MTP,such as pursuant to a PCR process. The module 114 (or any of the othermodules) can receive multiple MTPs so that the thermal cycle process canbe performed in parallel in order to increase input. The module 114 caninclude a device comprised of a centrifuge for spinning the MTPs beforeand after the thermal cycling to ensure all solutions in the MTPs areconcentrated at the bottom of each well and so are suitable for fluidichandling. As mentioned, the MTPs can be weighed prior and subsequent tothermal cycling to ensure that no evaporation or leakage has occurred.If a difference in weight of more than a certain threshold weight isdetected, the progress of that specific plate can be diverted to the endof the process line 100 and the user or tracking software notified.

[0105] With reference again to FIG. 1, the next module in the processline is the module 116, where a reagent, such as Shrimp AlkalinePhosphatase (SAP), is dispensed into the wells of the MTP. Prior todispensing the reagent, a workstation of the module 116 unseals thealuminum seal from the MTP to expose the wells of the MTP. The reagentis then added to all reaction wells in all plates, such as to destroyany unreacted nucleotide tri-phosphates in the wells. SAP is a commonreagent that can be dispensed using an array of solenoid valves linkedto a common reservoir of reagent which is temperature controlled formaximum shelf-life.

[0106] With reference to FIG. 1, the next module in the process line 100is the module 118, which is an incubation module. At the module 118, theMTP is subjected to an incubation process, such as for SAP incubation,if SAP was added at the previous module 116. The incubation module 118includes one or more workstations that facilitate the incubationprocess, such as a thermal cycling unit for SAP incubation andsubsequent heat inactivation. The module 118 can also include acentrifuge for spinning the MTP after thermal incubation of the MTP. Themodule 118 can also include a workstation that applies a seal to theMTP, such as a Polypropylene seal, that covers the wells of the MTP.

[0107] The next module is the module 120, which is the module where an“EXTEND” cocktail is added to the MTP, if required. The module 120includes a workstation comprised of a peeling unit, which removes thepolypropylene seal from the MTP. The module 120 can also include aworkstation comprised of a cooler that cools the MTP. In certainembodiments, the module 120 also includes a workstation comprised of asecond peeling unit for removal of the aluminum seal, if present, fromthe MTP. The module 120 can also include additional workstations, suchas a syringe array (such as a 384-syringe array) for rapid paralleltransfer of “EXTEND” cocktail from an “EXTEND” plate to a PCR reactionplate. The module 120 can also include a buffer position indicator foractive “EXTEND” plate if used for multiple PCR plates and a wash stationfor washing the MTP. A waste container can also be provided at themodule 120.

[0108] The next module is the module 122, which is a module where an“EXTEND” reaction is performed, as well as resin dispensing isperformed. An exemplary resin dispenser device is described in moredetail below. As in some of the previous modules, the module 122 caninclude workstations comprised of a centrifuge, a seal applicator forsealing the MTP, a thermal cycler for conducting an EXTEND reaction, anda peeling unit to remove polypropylene seal from the MTP after theEXTEND reaction.

[0109] With reference still to FIG. 1, the next module is the module124, where water addition, resin mixing, and chip printing is performed.The module 124 can include a workstation comprised of a water dispenserthat can be used to dispense water to the MTP. In one embodiment, a16-fold solenoid valve manifold is fed from temperature-controlledreservoirs to dispense water. The module 124 also includes a workstationcomprised of a centrifuge for spinning plates after water addition andprior to nanoliter transfer of samples from the MTP to a chip. Anin-line resin mixing station can also be deployed at the module 124, aswell as a device that dispenses samples from the MTP to a chip.

[0110] The computer system 101 controls the flow of plates from theconveyor line 110 into the module 124 to one of the work stations 124 a,124 b, 124 c and then back out to the conveyor line 110 again. Thetransfer of the plate from the conveyor line 110 to the module 124 canbe accomplished using a suitable transfer mechanism, such as atransverse conveyor belt that is oriented transverse to the direction ofthe conveyor line 110. When the plates encounter the transverse conveyorbelt, the plates are directed toward the module where appropriate.Similar control abilities are implemented by the computer system foreach of the other modules of the line 100.

[0111] Pin Alignment

[0112] As noted above, a sample delivery system constructed inaccordance with the invention aligns a pin array dispensing head totarget locations of a substrate, such as a chip, and automaticallydetermines any offset in pin alignment relative to the dispensing headfor successive blocks of pins. As used herein, “substrate” refers to aninsoluble support that can provide a surface on which or over which areaction may be conducted and/or a reaction product can be retained.Support can be fabricated from virtually any insoluble or solidmaterial. For example, silica gel, glass (e.g. controlled-pore glass(CPG)), nylon, Wang resin, Merrifield resin, Sephadex, Sepharose,cellulose, a metal surface (e.g. steel, gold, silver, aluminum, siliconand copper), a plastic material (e.g., polyethylene, polypropylene,polyamide, polyester, polyvinylidenedifluoride (PVDF)). Exemplarysubstrates include, but are not limited to flat supports such as glassfiber filters, glass surfaces, metal surfaces (steel, gold, silver,aluminum, copper and silicon), and plastic materials. The solid supportis in any desired form, including, but not limited to: a plate,membrane, wafer, a wafer with pits and other geometries and forms knownto those of skill in the art. Preferred support are flat surfacesdesigned to receive or link samples at discrete loci. Most preferred asflat surfaces with hydrophobic regions surrounding hydrophilic loci forreceiving, containing or binding a sample.

[0113]FIG. 4 shows an exemplary substrate comprising a chip 400 havingan array of target locations onto which sample materials will bedeposited during a printing process that takes place in the last module124. The chip illustrated in FIG. 4 includes three hundred eighty-fourtarget locations, arranged into a 16×24 grid. For easier and moreefficient handling, a group of chips can be collected together andplaced on a carrier tray. FIG. 5 shows a carrier tray 500 that mayaccommodate up to ten chips. The carrier tray 500 includes a pluralityof recessed chip holders 502 that can each receive a chip 400. The chipholders 502 are arranged into two rows, with five chip holders per row.

[0114]FIG. 6 shows a perspective view of the module 124 of the processline 100. As was discussed above, chip printing is performed at themodule 124. Thus, the module 124 includes at least one work stationcomprised of a delivery system or chip printing station 600. The chipprinting station 600 includes a movable dispensing head 604 thatincludes at least one pin array 606. Each pin array includes a pluralityof dispensing pins that can be dipped into the wells of an MTP so thatthe pins can aspirate material from the wells. The pins of the pin arraycan then be used to print the material onto a chip. In this regard, thechip printing station 600 also includes a loading station 616 wherechips can presented for loading of materials by the dispensing head 604.

[0115] The dispensing head illustrated in FIG. 6 includes sixteen arraysor blocks of pins, of which only the outermost 606 a, 606 b, 606 c, 606d, 606 e, 606 f, 606 g are visible in FIG. 6 (a reference to “606”without a letter suffix will be understood to be a reference to thecollection of all sixteen pin blocks generally, rather than to aparticular pin block). In one embodiment, each array contains a block oftwenty-four pins (in a 4×6 array), for a total of 384 pins in thedispensing head 604. It should be appreciated, however, that thequantity and spatial arrangement of the pins can vary. Each pin array606 can be removed from the dispensing head and replaced by areplacement array 606.

[0116] All of the sixteen pin arrays in the dispensing head 604 can bedipped into the MTP wells (such as a 384-well MTP) for aspirating samplematerial in the wells. The sample-loaded pins can dispense the samplematerial onto the chip one pin array at a time with the determined pitchin the MTP-to-chip reformatting process described below. In addition,less than all of the sixteen pin arrays 606 can be dipped into less thanall of the wells of the MTP for aspirating sample material from thewells. The pin arrays can then dispense sample material onto the chipone pin array at a time with the determined pitch or shift distance inthe MTP-to-chip dispensing and reformatting process. The aspiration ofthe remaining MTP wells can then be performed in order to complete thedispensing and reformatting process from the entire MTP wells onto thechip. The one-step sample material aspiration with multiple pin arrays,coupled with individual pin array printing onto the chip, can eliminatethe time-consuming steps for pin array washing, cleansing, and drying.Thus, the throughput of the process is maximized as a result.

[0117]FIG. 7 shows a perspective view of a single pin array 606. The pinarray 606 of FIG. 7 includes a plurality of dispensing pins 701, whereineach pin is configured to dispense a material in a well known manner.The pins 701 are mounted in a pin block 703. As mentioned, thedispensing pins are arranged in a rectangular array. In one embodiment,the array includes four rows of pins, with each row containing six pins.The pins 701 are positioned so that each pin can be aligned with acorresponding target location on a chip that is positioned below the pinarray.

[0118] With reference again to FIG. 6, the chip printing station 600 ispositioned adjacent a module conveyor line 301 for the module 124. Themodule conveyor line 301 is used to transport MTPs from the mainconveyor line 110 to the chip printing station 600. MTPs proceed alongthe main conveyor line 110 and, when appropriate, are directed into thechip printing station 600 by the computer system 101. The direction ofthe MTPs into the chip printing station may be accomplished, forexample, by utilizing the bar code of the MTP. The bar code can containinformation that directs the computer system 101 to forward a particularMTP to the chip printing station 600, such as when the MTP reaches themodule 124 as the MTP moves along the conveyor line 110. It should beunderstood that only one chip printing station is illustrated in FIG. 6for simplicity of presentation, and that the station illustrated in FIG.6 can include multiple stations.

[0119] With reference still to FIG. 6, the dispensing head 604 ismounted to a transport mechanism 614, such a track, that moves thedispensing head in a direction parallel to the module conveyor line 301and also perpendicular to the module conveyor line 301. Thus, thetransport mechanism 614 can be used to properly align the pins of thedispensing head 604 to the target locations of a chip onto whichmaterial will be printed. As described previously, alignment between thepins of the dispensing head 604 and the target locations of the chips isimportant for achieving accurate and valid testing results. Inaccordance with the invention, proper alignment is achieved with atwo-camera vision system that can identify and compensate for anymisalignment between the dispensing head and the chip, and between thepins and the dispensing head. The system is thereby unaffected by themisalignment that might otherwise occur, even after the dispensing headhas been aligned to a chip.

[0120] The vision alignment technique of the process line 100 involves achip alignment camera comprising a downward-looking camera 620 mountedto the side of the dispensing head 604 so that the downward-lookingcamera 620 is located in a fixed position relative to the dispensinghead 604, as shown in FIG. 6. The camera 620 is oriented so the camera620 can look down onto the top surface of a chip that is positionedbelow the camera 620. Thus, the target locations of the chip will be inthe camera field of view. The vision alignment technique also involves apin alignment camera comprising an upward-looking camera 622 that ismounted below the module conveyor line 301. The upward-looking camera622 is positioned so that it has a field of view that includes the pinsof the dispensing head 604. The downward-looking camera 620 ensuresthat, when the dispensing head is moved to a chip printing position, itis properly positioned above a chip for the pins with which it isinitially loaded and calibrated. As is conventional, a calibrationsequence is performed to ensure proper registration of the pins to thetarget locations at an initial process run. Thus, once the pin arrays606 are mounted to the dispensing head, there should be no concern ofmisalignment between the pins and chip target locations. In aconventional system, any change in pins presents an opportunity forpin-dispensing head misalignment to occur.

[0121] The present invention solves the problem of pin-dispensing headmisalignment by using the second camera 622 to check for any change inlocation of the pins relative to the dispensing head 604 whenever thepins are changed, such as when a pin array 606 is replaced. The secondcamera 622 looks up at the dispensing head 604 through a glass referenceplate 624 that is located in the field of view of the second camera 622.The pins of the dispensing head 605 are visible through a pin alignmentreticle on the glass reference plate 624 and in the field of view of thecamera 622. The position of a new block of pins on the dispensing head604 can be compared to the position of a prior block of pins, known tobe calibrated to delivery at the chip target locations, by noting anychange in pin position relative to the reticle, which is fixed relativeto the camera 622 and pins.

[0122]FIG. 8 shows a view down through the glass reference plate 624illustrated in FIG. 6, looking down at the upward-looking camera lens802 of the camera 622. In FIG. 8, the pin alignment reticle comprises aseries of “+” index marks 804, wherein one index mark is placed in eachcorner of the glass reference plate 624. Those skilled in the art willrecognize that many different index marks may be used as a reticle forpin alignment. All that is needed is to create a background patternagainst which the computer system may make a comparison of relative pinposition.

[0123]FIG. 9A shows a view from the perspective of the upward-lookingcamera 622, looking up through the reference plate 624 such that thereticle index marks “+” 804 are visible. The camera 622 also has a viewof the underside of a pin array 606 (the bottom tips of the pins 701 inthe pin array are represented as rectangles in FIG. 9A). When an arrayof pins is replaced by a new pin array, the position of the replaced pintips relative to the index marks 704 may be different from the positionof the previous array of pin tips relative to the index marks. Thecomputer system 101 can detect such a difference in position bycomparing a digital image of the original pin configuration with adigital image of the replacement pin array, as seen through thereference plate 624.

[0124] The computer system 101, when it detects a change in positionbetween a replaced array of pins and a new array of pins, may provide asignal to the operator and may halt operation of the chip printingstation 600, waiting for instruction or operator action. Alternatively,the computer system 101 can automatically identify and compensate forthe direction and magnitude of misalignment, through the aforementioneddigital image comparison technique. For example, the computer system 101can send instructions to the transport mechanism 615 to cause thetransport mechanism 615 to move the dispensing head 604 in order tocompensate for the misalignment.

[0125]FIG. 9B shows a view from the perspective of the downward-lookingcamera 620, looking down at a chip 400 that is positioned below thecamera and the dispensing head 604. The field of view of the downwardcamera includes chip alignment reticles 915 that are fixedly positionedin the field of view of the downward-looking camera 620. The reticles915 can be used to relatively locate index marks on the chip 400.Because the reticles 915 are fixedly located in the camera 620 field ofview and the camera 620 is fixedly located relative to the dispensinghead 604, the relative location between the chip index marks and thereticles 915 is an indication of the relative location between the chip400 and the dispensing head.

[0126] The flow diagram of FIG. 10 illustrates the operation sequence ofthe process line 100 in accordance with the two-camera alignmentchecking. In the first processing operation, represented by the flowdiagram box numbered 1002, the dispensing head 604 is moved to acalibration position. The exact location of this calibration positionwill depend on the particular installation of machinery at the chipprinting station 600, but will generally involve moving the dispensinghead to a known location above a particular chip of a chip tray that islocated at the loading station 616. Those skilled in the art willunderstand how to determine a suitable calibration position andprocedure for the system.

[0127] In the next operation, at block 1004 of FIG. 10, the dispensinghead 604 is moved so the upward-looking camera 624 can view the pins andcan locate a pin index position. This may comprise, as illustrated inFIG. 9, moving the dispensing head 604 so the pin arrays 606 provide acamera image in which the positions of the pin tips relative to the pinreticle in the camera field of view are substantially constant. Thoseskilled in the art will understand commonly used digital imageprocessing techniques that can be used to make comparison between thedigital images of the pin configurations, and will understand how toidentify misalignment.

[0128] At block 1005, the downward-looking camera 622 is used to locateindexing marks on a chip. The indexing marks may comprise any indiciathat appear in the camera field of view that may be useful in properpositioning (calibration) of the camera relative to the targetlocations. The index marks, for example, can comprise the targetlocations themselves. In one embodiment, the calibration image does notinvolve index marks that fill the camera field of view, but involves anedge of a chip. This provides a digital image that is more easilycompared for relative change from prior images, to more readily showsubtle changes in relative position.

[0129] At block 1008, the alignment of the pin arrays to the dispensinghead 604 and of the dispensing head 604 to the chips is determined fromthe upward and downward-looking cameras, respectively. The upwardlooking camera view is usually needed only when the pins or pin arraysare changed. It should not be necessary to perform the upward lookingpin calibration process during processing if there is no change in pinsor in the pin blocks, as it would be unlikely that the position of thepins relative to the dispensing head has changed. Preferably, thedownward-looking calibration will be utilized with every positioning ofthe dispensing head 604 over a chip for printing. If any camera viewindicates a misalignment, an affirmative outcome at the decision block1009, the computer system 101 will take corrective action. Amisalignment can be between the dispensing head and the chip or betweenthe pins of the dispensing head and the dispensing head. A misalignmentbetween the dispensing head and the chip is present where the relativelocations between the chip index marks and the chip alignment reticlehave changed between a current image and a previous image. Amisalignment between the pins and the dispensing head is present wherethe relative locations between the pins and the pin alignment reticlehave changed between a current image and a previous image.

[0130] The corrective action, indicated at block 1014, may comprisehalting operation of the process line, providing a message to theoperator, or automatically providing adjustment to operation, such as byadjusting the position of the pins or the dispensing head. For example,if the image from the downward-looking camera 620 indicates that thedispensing head is misaligned with respect to the original calibrationposition, then the dispensing head can be moved to re-align thedispensing head. If the image from the upward looking camera 622indicates that any of the pins are misaligned relative to the indexmarks 804, then the misaligned pins can be repositioned on thedispensing head. In any event, the corrective action to be taken willdepend on the needs of the particular process line installation. If nocorrective action is needed, then the system continues processing andprints sample material to a chip.

[0131] The downward camera 620 may be optionally used to check thevolume of sample material being deposited on the target locations. Toaccomplish this checking, after a chip has been printed, the dispensinghead 604 is moved to the downward-looking calibration position after achip has been printed, as indicated at block 1010 (which results from anegative outcome at block 1009). At the decision box numbered 1012, thecomputer system determines if the size of the sample spot on the chipfalls within a tolerance range for correct volumes of sample. If thesize of the spot indicates an incorrect volume, then at block 1014 thesystem takes corrective action.

[0132] The corrective action may comprise halting operation of theprocess line, or it may involve sending a message, or otherwise flaggingthe affected chip(s) for later disposal. In one embodiment, the computersystem 101 automatically checks the volume of dispensed material on thechip, determines if an adjustment to delivered volume should be made,and automatically makes the adjustment.

[0133] If the dispensed volume is within tolerance, an affirmativeoutcome at block 1012, then a calibration is performed at regularintervals of printing cycles, to ensure greater accuracy and operationwithin limits. The system checks (at block 1016) to determine if a pincalibration should be performed. Block 1016 indicates that the systemcomputer knows the interval at which calibration should be performed,and in one embodiment the system will query or prompt the systemoperator, or will automatically proceed with calibration at the propertime. Calibration is performed by returning to block 1002. If acalibration check of the pin relative to the dispensing head is notcalled for, a negative outcome at block 1016, then processing proceedswith normal processing of the next chip at the station, indicated atblock 1018, whereupon the dispensing head calibration to the next chipis performed at block 1006 and the other operations repeat.

[0134] It should be noted that other configurations of vision assistedalignment may be implemented without departing from the teachings of thepresent invention. For example, a single viewing camera may be utilized,in conjunction with mirror reflection, to perform the alignmentoperations described above. The FIG. 6 configuration, for example, maybe modified so that the reference plate 622 is replaced with a mirror,such that the only the downward-looking camera 620 is needed. When theupward view is required, observing the underside of the pin array, thedownward-looking camera will be positioned over the mirror referenceplate 622 to make an observation about the pin alignment. The reticlemarks of the reference plate will be printed on the top surface of themirror, so proper alignment checking may be performed. Thisconfiguration eliminates the need for two camera viewing systems.

[0135] Pin Array Reformatting

[0136] As noted above, a sample process line 100 constructed inaccordance with the present invention reformats a pin array of adispensing head to ensure that the spacing of the pin array at printingis reduced from the pitch at sample loading, preferably a multiple ofthe spacing of the target locations of a chip, in at least one dimension(reformatting in multiple dimensions may also be performed). In thesystem 100 illustrated in FIG. 1, the spacing of the pins at sampleloading time is an integral multiple of the wells. For example, atsample loading time, the MTP wells have a spacing of one well every 4.5mm, while the pin array block has a spacing of one pin every 9.0 mm.This initial spacing provides quick and efficient loading of the pins inthe wells. In accordance with the invention, the spacing of the pinarray within a block is then reduced at printing time to more nearlymatch the spacing of the chip target locations along two rows at a time.This reformatting of the pin array reduces the number of staggeredprinting actuations needed for the dispensing head. Thus, a greaternumber of pins may be arranged in a pin block, because the reduction inpin array pitch at printing permits more pins per actuation to beprinted to target locations. For example, with pair-wise reformatting ofall the rows of the pin array, the number of pins in a block can be fourtimes greater, and the number of staggered dispensing actuations can bereduced by one-fourth.

[0137]FIG. 11 is a side view of a pin array 1110 having at least one rowof pins 701 that are movably positioned. For comparison purposes, FIG.11 also shows a side view of an MTP 1112 below the pin array 110. TheMTP 1112 includes a plurality of wells 1113. FIG. 12 shows a top view ofthe pin array 1110, showing two rows of pins 701. The pin block 703includes a pitch changing comb 1112 that can engage protrusions 1202(shown in FIG. 12) on each of the pins 701. As described below, thepitch changing comb 1112 can be moved laterally (as exhibited by thedirectional arrow 1115 in FIG. 11) to reformat the pitch of the pins701. Thus, the pins 701 can be moved between a fully extended pitch(wherein the pin pitch is largest, as shown in FIGS. 11, 12) and afully-reduced pitch (wherein the pin pitch is smallest, as shown inFIGS. 13, 14).

[0138] In one embodiment, the pitch of the MTP wells is one well every4.5 mm, while the pitch of the pin array is one pin tip every 9.0 mm.Thus, as shown in FIG. 11, at the fully extended pitch, there is a pin701 aligned with every other well 1113 of the MTP 1112.

[0139]FIG. 13 is a side view of the pin array 1110 at the fully reducedpitch, from the same perspective as FIG. 9, while FIG. 12 is a top viewof the pin array 1110 at the fully reduced pitch, from the sameperspective as FIG. 10. In one embodiment, the pitch of the pin array1110 in FIG. 13 and FIG. 14 is one pin tip every 2.25 mm, which is areduced pitch from the fully extended configuration and is more nearlythe same pitch as the target locations on a chip. This permits thedispensing head 604 to be constructed with four times the number of pinsas before, because the reformatting permits more pins to be engaged inprinting at the same time. Reformatting from a pitch of 9.0 mm to 2.25mm (compare FIG. 11 and FIG. 12 with FIG. 13 and FIG. 14) permits thesame dispensing head blocks to be used with 384-well chips and also with96-well chips (a 96-well chip has target locations at a spacing of 2.25mm, a 384-well chip has a target location spacing of 1.125 mm). Thus,with reformatting from 9.0 mm to 2.25 mm, the number of staggeredprinting operations that are needed to print at the target locations isreduced by one-fourth.

[0140] As mentioned, each of the vertically oriented pins 701 has aprotrusion 1202 that engages a pitch changing comb 1112 that is movedlaterally when the reformatting is desired. FIG. 15 shows a group offour pins 701, with the protrusion 1202 of the end pin visible, as is aportion of the pitch changing comb 1112. Each row of pins whose pitch isto be changed has a corresponding pitch changing comb 1112. Thus, inFIG. 9, the side view shows a comb 1112 for the first row, and that combis 1112 also visible in the top view of FIG. 12. The second comb,referred to as comb 1112 a, for the second row is also visible in FIG.12. These same combs are visible in the corresponding reduced pitchdrawings of FIGS. 13 and 14.

[0141]FIG. 16 shows the sequence of reformatting as the pitch changingcomb 1112 is moved from the fully extended pitch to the fully reducedpitch. As the comb 1112 moves laterally, it engages each additional pin701 in the row, engaging a new pin as the comb moves along from right toleft in the drawing. In FIG. 16, each instance of engaging a new pin 701is indicated as a step of the reformatting operation, which emphasizesthe stepped appearance of the engaging surface of the comb 1112. In thefirst step, Step 1, the comb 1112 is shaded to highlight its positionfor easier understanding of the operation. At each illustrated step ofFIG. 16, a pin protrusion is indicated as a solid black square, again tohighlight its position for easier understanding.

[0142] Thus, at Step 1, the top most pin protrusion is already engagedwith the highest step of the comb 1112. At Step 2, the comb 1112 hasmoved toward the left and the next highest step of the comb 1112 hasengaged the next highest protrusion, which is located on the next pin.The first pin remains engaged with the comb 1112, and is moved along bythe comb 1112 so that its spacing from the second pin is now reduced.Both the first pin and the second pin are moved together toward thethird pin and the spacing from the third pin to the second pin and firstpin is reduced. In the third step, the comb 1112 has engaged the thirdpin. Now these three pins are moved along, and the process continuesuntil all twelve pins in the pin block are moved. At the last step (step12), all twelve pins have been moved and have a new uniform pitch thatis one-quarter of its prior pitch, being more nearly the same pitch asthe target locations on the chip.

[0143] It should be appreciated that the pitch of the pins in each pinblock 606 can be reformatted independently of every other pin block onthe dispensing head. For example, the pins of pin block 606 a can be setto a first pitch and the pins of pin block 606 b can be set to adifferent pitch than the pins of block 606 a. Thus, the pitch of eachpin block 606 can be formatted independently of the other pin blocks, orall of the pin blocks 606 can be formatted as a common group. The pinblock 606 a can be set to a first pitch suitable for aspirating from anMTP, and then set to a second pitch suitable for dispensing to thetarget locations on a chip, while the pin block 606 b (or any other pinblock) can be set to a different pitch during this process. This enablesa higher throughput of MTP processing than if the pin blocks all had tobe set to a common pitch.

[0144] It should be appreciated that the pitch of the pin array may bereduced to be more nearly equal to the pitch of the target locations onthe chip, the limitation being the diameter of the pins themselves. Thatis, the pins of the preferred embodiment have a diameter (including anyspring actuation or support structures) that precludes a spacing that isidentical to that of the target locations. Those skilled in the art,however, will understand that the technique described herein may be usedto reformat the pins to a pitch that is the same as the targetlocations.

[0145] Resin Particle Dispensing

[0146]FIG. 17A is a perspective view of the resin dispensing module 122(FIG. 1) of the processing line. Microtiter plates proceed along themain conveyor line 110 and, when appropriate, are directed onto theconveyor 1502 of the resin dispensing module 122 by the computing system101. It should be understood that only one resin dispensing station isillustrated in FIG. 17A for simplicity of presentation, and that theresin dispensing module 122 illustrated in FIG. 1 includes multiplestations, each of which performs the resin dispensing task.

[0147] The resin dispensing module 122 includes a conveyor 1502, whichdirects the MTPs 1504 to the module 122. It also includes a resindispensing assembly 1508, which is made up of a number of hollow tubes1802 (shown in more detail in FIGS. 20A-C). The hollow tubes 1802 can bemolded, welded, mechanically attached (such as by individually threadingthem), or otherwise attached to an array plate 1601, as shown in moredetail in FIGS. 18A and 18B. The resin dispensing assembly 1508 ismounted on a transport mechanism 1510. The transport mechanism 1510includes a guide rail 1511 along which the dispensing assembly 1508slides. The guide rail 1511 includes sensors 1520 and 1522 at itsproximal and distal ends respectively, and these sensors are used todetect the position of the dispensing assembly 1508. The dispensingassembly 1508 can be moved, for example, pneumatically or hydraulicallyalong the guide rail 1511. The module 122 also includes a resinreservoir assembly 1535 and a skimming plate 1530, each of which will bediscussed in more detail below.

[0148]FIG. 17A shows an 1504 that has been directed from the mainconveyor line 110 onto the resin dispensing line 1502. In FIG. 17A, thehollow tube array 1506 has been loaded with resin particles and ispositioned over the MTP 1504, ready to dispense resin particles fromeach of the hollow tubes 1802 into the wells of the MTP 1504. The hollowtubes 1802 of the array 1506 are suspended from the dispensing assembly1508 that is mounted to the transport mechanism 1510 that moves thedispensing assembly in a direction perpendicular to the module line 1502along a Y axis. Positioned underneath the MTP 1504 is a lifting platform1555, which aligns the MTP 1504 with the array 1506, and lifts the MTP1504 slightly toward the array 1506.

[0149] The dispensing assembly 1508 starts from a point of origin justabove the conveyor 1502 and the lifting platform 1555, at the proximalend of the guide rail 1511. A sensor 1520 (see FIG. 17C) attached to theguide rail 1511 is used to detect the position of the dispensingassembly 1508. From that point of origin, the dispensing assembly 1508is moved distally along the guide rail 1511 (along the Y axis), until itstops at the distal end of the guide rail 1511, where a sensor 1522 (seeFIG. 17C) is stationed to detect the arrival of the dispensing assembly1508. Looking now at FIG. 17C for a view of the module 122 from therear, the dispensing assembly 1508 stops above a skimming plate 1530.FIG. 17D is a side section view of the resin dispensing module describedabove.

[0150] The skimming plate 1530 can be made of any durable and stiffmaterial, and in one embodiment is made of machined aluminum with astainless steel perimeter. The skimming plate 1530 has holes throughwhich the hollow tubes of the array 1506 slide. The skimming plate 1530can have at least as many holes as there are hollow tubes on the array1506, but not fewer. In one embodiment, the array 1506 has 384 hollowtubes, and the skimming plate 1530 has 384 holes. In another embodiment,the array 1506 has 96 hollow tubes, and the skimming plate has 96 holes.In yet another embodiment, the array 1506 has 1,536 hollow tubes and theskimming plate has 1,536 holes in it. The holes of the skimming plate1530 are aligned with the hollow tubes of the array 1506 so that all ofthe hollow tubes will slide simultaneously through each of theircorresponding holes when the dispensing assembly 1508 is positioned overthe skimming plate 1530.

[0151] Either before, during, or after the dispensing assembly 1508 ispositioned over the skimming plate 1530, the resin reservoir 1540 isdeployed. The resin reservoir assembly 1535 deploys the resin reservoir1540, which can be pneumatically or hydraulically guided along the Xaxis toward the skimming plate 1530. It comes to a stop just under theskimming plate 1530.

[0152] Once the resin reservoir 1540 is in position underneath theskimming plate 1530, and the array 1506 is in position over the skimmingplate 1530, the array 1506 is pneumatically or hydraulically loweredalong the Z axis. Vertical displacement shafts 1630 on the dispensingassembly 1508 slide vertically into vertical displacement bores 1632,thus allowing the array 1506 to drop vertically. This allows the hollowtubes 1802 to slide through the holes of the skimming plate 1530, andinto the resin reservoir 1540, filling the distal ends of the tubes 1802with resin. The force of lowering the array 1506 into the reservoir 1540pushes resin particles up into each of the hollow tubes 1802. Thefriction between particles after they have been pushed into the tubes1802 holds the particles within the tubes as the array 1506 is moved outof the resin reservoir 1540. The resin particles also becomefrictionally engaged with the inner surfaces of the hollow tubes 1802(as shown in more detail in FIG. 20C).

[0153] The array 1506 is then pneumatically or hydraulically raisedalong the Z axis, and the hollow tubes 1802 are withdrawn from the resinreservoir 1540 and are raised through the holes of the skimming plate1530. The diameter of each of the holes in the skimming plate 1530 isjust slightly larger than the diameter of each of the hollow tubes 1802,such that when the hollow tubes 1802 are withdrawn through the holes ofthe skimming plate 1530, the outside surfaces of the hollow tubes 1802are skimmed clean by the skimming plate 1530. This ensures that unwantedamounts of resin do not cling to the outside surface of the hollow tubesand become inadvertently dispensed into an MTP 1504.

[0154] In an alternative embodiment, the array 1506 can remain staticwhile the resin reservoir 1540 is raised to meet the array 1506. Thereservoir can engage the skimming plate 1530 and raise it toward thearray 1506, resulting in the hollow tubes 1802 being threaded throughthe holes in the skimming plate 1530. Once the hollow tubes are filledwith resin, the reservoir 1540 can be lowered along with the skimmingplate.

[0155] Once the array 1506 is completely withdrawn vertically, thedispensing assembly 1508 is pneumatically or hydraulically guided alongthe Y axis back to its point of origin. Either before, during, or afterthe dispensing assembly 1508 arrives at its point of origin, an MTP 1504will be guided along the conveyor 1502 and will come to a rest above thelifting platform 1555 and just underneath the array 1506.

[0156] The lifting platform 1555 is stationed at a predeterminedposition beneath the point of origin of the dispensing assembly 1508.When the MTP 1504 slides over the lifting platform 1555, the liftingplatform is raised upward and catches the MTP 1504. The lifting platformcan have raised edges that fit snugly around the MTP 1504, thus aligningthe MTP 1504 with the array 1506, which is above it. Alternatively, thelifting platform 1555 can have other means of aligning the MTP 1504 withthe array 1506. For example, the lifting platform 1555 can have magnetson its upper surface with corresponding metal points on the bottomsurface of the MTP 1504, or the metal points and magnets can be reversedso that the magnets are on the MTP 1504, while the metal points are onthe lifting platform 1555. In another embodiment, the upper surface ofthe lifting platform 1555 can have one or more holes, bores, cavities,grooves, or slots into which corresponding protuberances on the bottomsurface of the MTP 1504 fit, or vice versa.

[0157] The MTP 1504 can have a number of wells equal to the number ofhollow tubes 1802. The wells of the MTP 1504 and the hollow tubes 1802in the array 1506 will be aligned, and the array will be pneumaticallyor hydraulically lowered along the Z axis toward the MTP 1504. The array1506 will then come to a rest and plungers 1804 within each of thehollow tubes 1802 will be lowered, causing the resin to be pushed out ofthe hollow tubes and into the wells of the MTP 1504.

[0158] Meanwhile, the resin reservoir 1540 can be pneumatically orhydraulically guided back to its point of origin, where it can slideunderneath a compacting lid 1745, which engages the top of the reservoir1540. A compactor can pneumatically or hydraulically press down againstthe lid 1745 to pack the resin so that a flat and uniform resin bed isachieved. In addition, a vibrator 1765 (as shown in FIG. 19B) can beused to vibrate the compacting lid 1745 to further pack the resinparticles into a flat and uniform bed.

[0159] In the preferred embodiment, the number of hollow tubes in thearray 1506 is equal to the number of wells in the MTP 1504. Thus,loading of all hollow tubes takes place simultaneously, and dispensingof all hollow tubes takes place simultaneously, and loading of allmicrotiter wells occurs simultaneously. The resin dispensing module ofthe present invention thereby assists in throughput increasing efforts.

[0160] The Resin Dispensing Assembly FIG. 18A is a closer view of theresin dispensing assembly 1508. The resin dispensing assembly includesan array 1506 of hollow tubes 1802. The hollow tubes 1802 can be welded,integrally molded, or mechanically attached (such as by individuallythreading) to a rectangular array plate 1601, having a length L, a widthW, and a depth D.

[0161] In one embodiment the array plate 1601 is solid, and a number ofholes are bored through it from its top surface 1611 to its bottomsurface 1612. The number of holes is equal to the number of hollow tubes1802 in the array 1506. The hollow tubes 1802 can be attached to thebottom surface 1612 of the array plate 1601 in any manner known to thosein the art, such as welding or securing with an adhesive. The hollowtubes and the bored holes can all be aligned with one another and canhave the same diameters, so that the inner walls of the bored holes lineup exactly with the inner walls of the hollow tubes. For example, in anarray with 384 hollow tubes 1802, this results in an array plate 1601with 384 passages leading from 384 holes on its top surface through 384hollow tubes 1802 and out the distal openings 1803 (as seen in FIG. 20A)of those 384 hollow tubes 1802.

[0162] In another embodiment, the array plate 1601 can have a number ofbored holes leading from openings in the top surface 1611 of the plateto openings on the bottom surface 1612 of the plate. The number of boredholes can be equal to the number of hollow tubes 1802. The hollow tubes1802 can be radially sized to fit coaxially within the bored holes, andthe proximal ends thereof can be inserted through the openings on thebottom surface 1612 of the plate. The hollow tubes 1802 can then beforced through the bored holes until the proximal ends of the hollowtubes 1802 are flush with the top surface 1611 of the plate. The hollowtubes can be coaxially engaged with the bored holes through frictionalengagement, by an adhesive, or by any other means known to those withskill in the art. In any case, the result is an array 1506 of hollowtubes 1802, the hollow tubes protruding form the bottom surface 1612 ofan array plate 1601 having a corresponding array of bored holes.

[0163] The array plate 1601 is connected to an upper plate 1603 by twovertical support walls 1602. The array plate 1601 can be bolted orwelded to the vertical support walls 1602, which can be bolted or weldedto the upper plate 1603. Isolated from any vertical force exerted oneither the upper 1603 or array plate 1601 and floating in between thetwo is a plunger plate 1605. The plunger plate 1605 can float on one ormore springs placed in between the top surface 1611 of the array plate1601 and the bottom surface 1614 of the plunger plate 1605. The devicealso has at least two stop posts 1610. The stop posts 1610 includeflanged terminals 1616 that prevent the plunger plate 1605 from floatingbeyond a predetermined distance above the array plate 1601. The stopposts 1610 also align the plunger plate 1605 and array plate 1601. Inaddition, the stop posts 1610 can have springs (not shown) fittedcoaxially around them in between the array plate 1601 and plunger plate1605. These stop post springs can be used in lieu of or in addition tothe springs discussed above.

[0164] Protruding from the bottom surface of the plunger plate 1605 area number of plungers 1804 (as shown in more detail in FIG. 18B). Thenumber of plungers 1804 can be equal to the number of hollow tubes 1802in the array 1506. The plungers 1804 are aligned with the holes on thetop surface of the array plate 1601, and they are inserted into thehollow tubes 1802 through those holes. The plungers 1804 are at least aslong as the hollow tubes 1802. The plungers 1804 are used tosimultaneously push the resin out of each of the hollow tubes 1802.

[0165] The amount of resin that is collected by the hollow tubes dependson how much space there is between the bottom of the plungers 1807 andthe bottom of the hollow tubes 1803 (as shown in more detail in FIGS.20A and 20C). This space can be controlled by adjusting the verticalposition of the plungers 1804 within the hollow tubes 1802. Thisadjustment is made using an adjustment screw 1615. The adjustment screw1615 can be threaded through a threaded hole in the upper plate 1603 andextend out through a corresponding bottom hole. The end of the screw1615 can be used as a stopper against the upward force of the plungerplate 1605 caused by the springs. The screw 1615 can be calibrated anddemarcated so that the amount of resin desired for a particular assaycan be adjusted quickly and easily using the screw.

[0166] The plungers 1804 can be forced down using a compressing assembly1625, which can be placed on top of the upper plate 1603, and joined tothe top of the plunger plate 1605 through the upper plate 1603. Thecompressing assembly 1625 can be pneumatic or hydraulic, and like all ofthe other pneumatic or hydraulic components of the system, can becomputer controlled. The dispenser assembly 1508 thus allows forcontrolled delivery of resin or other chemical or biological reagents.

[0167] The Hollow Tubes

[0168]FIG. 20A is a schematic representation of the lower portion of onehollow tube 1802. A solid plunger 1804 moves up and down within thehollow tube 1802, and is shown in FIG. 20A in its most upward position.At this raised position, it should be apparent that the volume of resinparticles that will be picked up in the tube is defined by the internaltube volume from the bottom 1807 of the plunger 1804 to the open end1803 of the hollow tube 1802, represented by the portion designated bythe brackets 1806. After the hollow tube array is lowered toward the MTP1504 and is in position over the MTP wells, the plungers 1804 will belowered, so they push out all the contents (resin particles) containedin the tube 1802, out and into a corresponding well of the MTP 1504.This is illustrated in FIG. 20B, which depicts the plunger 1804 pusheddown to its farthest downward location. Alternatively, the hollow tube1802 can be raised and moved upward in relation to the plunger 1804rather than the plunger 1804 being lowered. In any case, the plunger1804 pushes the resin particles 1820 out of the space 1806.

[0169] As noted above, the system 100 moves the plungers 1804 down allof the hollow tubes simultaneously. As explained, this may beimplemented with a flat plunger plate 1605 connected to all of theplungers 1804, thereby exerting a force simultaneously on all theplungers 1804 and moving them in unison. Thus, as shown in FIGS. 18A,18B, and 20C, the top surface 1808 of the plungers will preferably beconnected to a solid plunger plate 1605.

[0170] The plungers 1804 can include one or more channels formedcoaxially around the outer surface of their distal ends. For example,FIG. 20C shows a plunger 1804 with a channel formed coaxially on theoutside surface at its distal end. An O-ring 1819 can be coaxiallymounted into the channel. The O-ring 1819 seals the outer surface of theplunger 1804 against the inside surface of the hollow 1802.Alternatively, a bushing can be coaxially mounted over the plungers1804, to seal the plungers 1804 against the inside surface of the hollowtubes 1802.

[0171] Although the hollow tube is discussed herein with respect to theobjective of collecting, transporting, and dispensing resin particles,it should be understood that the device can be used to collect,transport, and dispense any solid material, such as any type ofbiological or chemical reagent.

[0172] The Particle Reservoir Assembly

[0173]FIGS. 17A, 17B, and 17C show that the resin dispensing module 122also includes a resin reservoir assembly 1535, which includes a resinreservoir 1540 where resin, or some other biological or chemicalreagent, can be stored for acquisition by the array 1506. FIGS. 17B and19B show the resin reservoir 1540 in its resting state. As shown in moredetail in FIG. 19A, the resin reservoir 1540 can include a foundation1720 with a number of springs 1715 attached to it. The springs cansurround a reservoir base 1717, which rests on top of the foundation1720. The reservoir base 1717 can include one, two, three, or moreO-rings 1719 placed in horizontal channels encircling the base. Areservoir collar 1710 is placed on top of the springs. The reservoircollar 1710 can be any shape, but it must coincide with the shape of thebase 1717. If the base 1717 is cylindrical, then the collar 1710 must beshaped in the form of a hollow cylinder. If the base 1717 is rectangular(as shown), then the collar 1710 must have a rectangular opening sizedto receive the base 1717. Lengthwise, the top of the collar includesgrooves 1728 that are used to secure the compacting lid 1745 against thecollar 1710.

[0174] The top of the compacting lid 1745 is flat and is connected to acompressor 1525, which can be pneumatically or hydraulically operated.The lid 1745 includes a hollowed out portion, and a vibrator plate 1765is inserted into it. The rear end of the vibrator plate 1765 includes astem that is connected to a pneumatic or hydraulic vibrator (not shown)for vibrating the plate. Alternatively, the vibrator plate 1765 caninclude internal vibrating components and an internal power source.Thus, when the vibrator plate 1765 vibrates, it causes the entire lid1745 to vibrate. The underside of the compacting lid 1745 is concave andhas a channel with an O-ring to seal the lid 1745 against the collar1710. The underside surface may be coated with a stick-resistantmaterial, such as “Teflon” or the like. Depending on the particlematerial, other treatments might be desirable for ensuring propercompacting and presenting a uniform surface to the tube array, includingelectrical charge or airflow.

[0175] The reservoir 1540 is formed when the base 1717 is insertedthrough the collar 1710, the base 1717 forming the bottom of thereservoir, while the collar 1710 forms the walls.

[0176] In operation, the foundation 1720, which can be mounted on tracks1722, can slide underneath the skimming plate 1530. The skimming plate1530 can be detached and moved out of the way so that the operator canload the reservoir 1540 with resin or some other biological or chemicalreagent. Once the reservoir 1540 is loaded, the foundation 1720 canslide pneumatically or hydraulically back to its point of originunderneath the compacting lid 1745. It may be advantageous to compactthe particles that are in the reservoir 1540. To accomplish that, thecompressor 1525 pushes down on the lid 1745, which is forced onto thecollar 1710 and pushes down on it. The collar 1710, which rests onsprings 1715, is consequently forced downward over the base 1717 andtoward the foundation 1720 until the underside of the lid 1745 comesinto contact with the resin in the reservoir 1540. The amount ofpressure required will depend on the composition of the resin particles,as will be known to those skilled in the art. Meanwhile, the vibratorplate 1765 causes the lid 1745 to vibrate. The vibration causes thecompacting lid 1745 to further pack the resin particles into a flat anduniform bed. Alternatively, a pneumatic or hydraulic vibrator can beconnected to the collar 1710, base 1717, or foundation 1720, and canshake or vibrate any of those structures.

[0177] Once compaction is complete, the compressor 1525 decompresses,causing a pause in the downward force. Without the extra downward force,the springs 1715 push the collar 1710 and lid 1745 back upward, and theresin in the reservoir 1540 is ready for a new cycle of resindispensing.

[0178] In an alternative embodiment, the foundation 1720 may bepneumatically or hydraulically raised to force the resin against the lid1745, rather than forcing the lid downward. In either case, the effectis to force the underside of the lid against the resin, thus compactingthe resin.

[0179] The resin compacting protocol can be repeated several times untilthe resin is sufficiently compacted and ready for a cycle of dispensing.The compacting lid 1745 is useful because, as the hollow tubes 1802 arewithdrawn from the reservoir 1540 in their loaded state, they may likelyleave a corresponding array of voids in the particle bed of thereservoir 1540, corresponding to the volumes that were drawn out of thereservoir 1540 and pushed into the hollow tubes 1802. Therefore, the lid1745 is used to rearrange the particles and provide a substantiallyuniform bed of resin particles. This ensures that a level surface willbe presented to the tube array at the next loading cycle of thedispensing module.

[0180] Computer Control

[0181] The process line 100 illustrated in FIG. 1, whose operation hasbeen described above in conjunction with the flow control,reconfiguration, alignment, and reformatting operations, preferably iscontrolled by the computer system illustrated in FIG. 1. That computersystem includes a conventional programmable computer, and communicateswith the devices of the various process line stations over a datanetwork, to thereby control the operations that occur at each module andeach station. An exemplary computer embodiment for performing thesecontrol functions is illustrated and described below.

[0182]FIG. 21 is a block diagram of a computer that may be used toimplement the process line control described herein. It should beunderstood that the process line control functions described herein maybe performed with a single computer, or may be used in conjunction withone or more computers that may communicate with each other over anetwork to share data. Those skilled in the art will appreciate that thevarious processes described above may be implemented with one or morecomputers, all of which may have a similar computer construction to thatillustrated in FIG. 21, or may have alternative constructions consistentwith the capabilities described herein.

[0183]FIG. 21 shows an exemplary computer 2000 such as might compriseone of the computers that implements the functions and actions describedabove. Each computer 2000 operates under control of a central processorunit (CPU) 2002, such as a “Pentium” class microprocessor and associatedintegrated circuit chips, available from Intel Corporation of SantaClara, Calif., USA. A computer user can input commands and data from akeyboard and computer mouse 2004, and can view inputs and computeroutput at a display 2006. The display is typically a video monitor orflat panel display. The computer 2000 also includes a direct accessstorage device (DASD) 2008, such as a hard disk drive. The memory 2010typically comprises volatile semiconductor random access memory (RAM).Each computer preferably includes a program product reader 2012 thataccepts a program product storage device 2014, from which the programproduct reader can read data (and to which it can optionally writedata). The program product reader can comprise, for example, a diskdrive, and the program product storage device can comprise removablestorage media such as a magnetic floppy disk, a CD-R disc, a CD-RW disc,or DVD disc.

[0184] The computer 2000 can communicate with other computers and withthe devices of the process line over a computer network 2016 (such as alocal area network, or the Internet or an intranet) through a networkinterface 2018 that enables communication over a connection 2020 betweenthe network 2016 and the computer 2000. The network interface 2018typically comprises, for example, a Network Interface Card (NIC) or amodem that permits communications over a variety of networks.

[0185] The CPU 2002 operates under control of programming steps that aretemporarily stored in the memory 2010 of the computer 2000. When theprogramming steps are executed, the computer performs its functions.Thus, the programming steps implement the functionality of the processline control system described above. The programming steps can bereceived from the DASD 2008, through the program product storage device2014, or through the network connection 2020. The program productstorage drive 2012 can receive a program product 2014, read programmingsteps recorded thereon, and transfer the programming steps into thememory 2010 for execution by the CPU 2002. As noted above, the programproduct storage device can comprise any one of multiple removable mediahaving recorded computer-readable instructions, including magneticfloppy disks and CD-ROM storage discs. Other suitable program productstorage devices can include magnetic, tape and semiconductor memorychips. In this way, the processing steps necessary for operation inaccordance with the invention can be embodied on a program product.

[0186] Alternatively, the program steps can be received into theoperating memory 2010 over the network 2016. In the network method, thecomputer receives data including program steps into the memory 2010through the network interface 2018 after network communication has beenestablished over the network connection 2020 by well-known methods thatwill be understood by those skilled in the art without furtherexplanation. The program steps are then executed by the CPU 2002 therebycomprising a computer process. If desired, updates to the computersoftware may be achieved in this manner. FIG. 21 shows a device 2022connected to the network 2016 in a similar configuration as the computer2000. It should be apparent that the device 2022 may comprise anothercomputer and may also include one or more of the devices comprising theprocess line 100, as described above.

[0187] Thermal Cycling

[0188] As noted above, some systems make use of thermal cyclingoperations to subject the materials to temperature regimens. Theautomated process line illustrated in FIG. 1, constructed in accordancewith one embodiment of the present invention, introduces fluids ofdifferent temperatures to a configuration of multiple flow pathwaysformed by flow cell assemblies on which microtiter plates (MTPs) aremounted and fixed by upper heated lids. Fluids of different temperaturesare supplied from fluid reservoirs to the underside of the microtiterplate. Valves switch fluid from a selected reservoir to a manifold thatdistributes the fluid stream to the individual flow cells. Theunselected reservoirs remain in continuous circulation by bypassing themanifold to maintain the system at a fixed bath temperature.

[0189] In accordance with the invention, an insert is integrated intoeach flow cell assembly, such that the insert supports the wells of theMTP from beneath and contains flow directing guide elements that promotea uniform fluid pressure over the whole length of the MTP perpendicularto the direction of flow. This ensures a uniform flow over the wells ofthe MTP. The insert provides faster temperature change of the wellcontents and provides a more uniform distribution of temperature throughall the wells of the plate and within each of the wells. The flowdirecting guide elements, and selection of an appropriate flow rateprovide a uniform temperature distribution across the active flow cellarea. Upon completion of the thermal cycling process, the MTPs are driedand brought to ambient temperature by introducing compressed gas.

[0190]FIG. 22 shows a schematic, top view diagram of the process linemodule 114, where thermal cycling of one or more MTPs can be performed.The module 114 includes a workstation comprised of a thermal cyclingsystem 2100 that includes one or more thermal cycling stations 2105,including stations 2105 a, 2105 b, 2105 c, 2105 d, 2105 e. Throughoutthis description various items are referred to generally andcollectively using a reference numeral, and sometimes referred toindividually using a reference numeral followed by a letter suffix. Itshould be appreciated that items that are referred to using a commonreference numeral are identical in structure unless otherwise noted.FIG. 23 shows five thermal cycling stations 2105, although it should beappreciated that the thermal cycling system 2100 can include any numberof stations.

[0191] As shown in FIG. 22, the thermal cycling system 2100 is locatedadjacent the module conveyor line 301 of the module 114. An MTP can betransported by the module conveyor line 301 to each of the thermalcycling stations 2105 for loading onto the thermal cycling stations.Each thermal cycling station 2105 is configured to receive a single MTP,such as via a conveyor belt that transfers an MTP from the moduleprocess line 301 to each station 2105.

[0192]FIG. 23 is a schematic side view of the thermal cycling system2100, showing various components of the thermal cycling system 2100.Each station 2105 is configured to hold a microtiter plate assembly2110, which includes a microtiter plate that has been loaded onto thestation 2105 and various other components that are used to thermallycycle the microtiter plate, as described more fully below. The thermalcycling system 2100 further includes one or more fluid reservoirs 2115that each contain a fluid that can be distributed to the microtiterplate assemblies 2110. In this regard, each reservoir includes an inletpipe 2120 through which fluid can flow into the respective reservoir2115, and an outlet pipe 2125 through which fluid can flow out of therespective reservoir 2115. The inlet pipe 2120 and outlet pipe 2125 ofeach reservoir 2115 connects to a manifold and valve system 2130 thatpermits an operator to selectively flow fluids from any of thereservoirs 2115 to any of the microtiter plate assemblies 2110. Each ofthe stations 2105 includes a corresponding inlet pipe 2135 through whichfluid from the manifold and valve system 2130 can be flowed into amicrotiter plate assembly 2110, as well as an outlet pipe 2140 throughwhich fluid can be flowed out of a microtiter plate assembly 2110 to themanifold and valve system 2130.

[0193] Each of the reservoirs 2115 is temperature controlled in awell-known manner so that the fluid in each reservoir can be maintainedat a predetermined temperature. In FIG. 23, the reservoir 2115 a is at atemperature T1, the reservoir 2115 b is at a temperature T2, and thereservoir 2115 c is at a to temperature T3. It should be appreciatedthat the thermal cycler system can include more or less reservoirs thanwhat is shown in FIG. 23.

[0194] As shown in FIG. 23, a temperature controlled plate 2240 islocated above the microtiter plate assemblies 2110. The plate 2240 canbe moved upward and downward relative to the microtiter plate assemblies2110, such as by a pneumatic lift 2245 that is attached to the plate2240. The plate 2240 can move downward toward the assemblies 2110 sothat the plate contacts the assemblies 2110 and transfers heat to theassemblies 2110. In this manner, the assemblies 2110 can be heated to adesired temperature.

[0195]FIG. 24 is a schematic side view of the microtiter plate assembly2110, which shows the flow path through which fluid can flow through themicrotiter plate assembly 2110 during thermal cycling. FIG. 24 omitsstructural details of the microtiter plate assembly 2110, whichstructural details are shown and described in other figures below. Themicrotiter plate assembly 2110 includes a microtiter plate 2310 that isremovably positioned atop a flow cell assembly 2315. The flow cellassembly 2315 guides fluid through a flow path so that the fluidcontacts at least a portion of the microtiter plate 2310, such as abottom surface of the microtiter plate 2310. As described in detailbelow, the fluid is guided in such a manner that it flows evenly acrosseach of the wells of the microtiter plate 2310. FIG. 24 shows thegeneral direction of the flow path using a collection of arrows.

[0196] The flow cell assembly 2315 includes three fluid flow regionsthat collectively guide fluid through the flow path. The fluid flowregions include an inlet/outlet flow region 2320, an intermediary flowregion, 2335, and a thermal cycling flow region 2345. The inlet/outletflow region 2320 is the portion of the flow cell assembly 2315 throughwhich fluid flows into the flow cell assembly 2315 from a respectiveinlet pipe 2135 (shown in FIG. 23) and through which fluid flows out ofthe flow cell assembly 2315 through a respective outlet pipe 2140 (shownin FIG. 23). The inlet/outlet flow region 2320 includes an inlet conduit2325 through which fluid flows into the flow cell assembly 2315, as wellas an outlet conduit 2330 through which fluid flows out of the flow cellassembly 2315. In an exemplary configuration, the inlet conduit 2325guides the fluid so that it flows in a substantially upward directioninto the flow cell assembly 2315 from the inlet pipe 2125 (shown in FIG.23), and the outlet conduit 2330 guides the fluid in a substantiallydownward direction out of the flow cell assembly 2315 into the outletpipe 2120 (shown in FIG. 23).

[0197] The flow cell assembly 2315 further includes the intermediaryflow region 2335 in which (1) fluid is guided from the inlet/outlet flowregion 2320 to an inlet opening 2340 that leads to the thermal cycleflow region 2345; and (2) fluid is guided from an outlet opening 2342(that leads from the thermal cycle flow region 2345) to the inletconduit 2325 of the inlet/outlet flow region 2320. As described in moredetail below, the intermediary flow region 2335 includes one or moreflow guide members, such as baffles, that guide fluid through theintermediary flow region 2335 in a predetermined manner toward a desiredtarget location. In one embodiment, the fluid in the intermediary flowregion 2335 flows in a sideways, or horizontal, direction as it travelsfrom the inlet conduit 2325 to the inlet opening 2340 and from theoutlet opening 2342 to the outlet conduit 2325.

[0198] As shown in FIG. 24, the flow cell assembly 2315 further includesthe thermal cycling flow region 2345, in which fluid flows in contactwith the microtiter plate 2310 to absorb heat from the microtiter plate2310. The thermal cycling region 2345 includes flow guides that formflow channels through which fluid flows in a predetermined flow patternunderneath rows of wells of the microtiter plate, as described in moredetail below. The fluid enters the thermal cycling region 2345 from theintermediary flow region 2335 through the inlet opening 2340 and exitsthe thermal cycling region 2345 to the intermediary flow region 2335through the outlet opening 2342.

[0199]FIG. 25 shows an exemplary microtiter plate 2310, which includesone or more wells 2415. For clarity of illustration, only one of thewells 2415 is labeled with a reference number. The wells 2415 can bearranged in a series of rows and columns to form an array of wells 2415.Those skilled in the art will appreciate that the microtiter plate 2310can have any number of wells that are arranged in any number of rows andcolumns. For example, some microtiter plates, such as the microtiterplate 2310 of FIG. 25, have twenty-four wells arranged in a six row byfour column array, and other microtiter plates have ninety-six wellsarranged in a twelve row by eight column array. Another conventionaltype of microtiter plate includes three hundred eighty-four wellsarranged in a 16×24 array. The wells can be arranged in any variety ofrow and column configurations.

[0200]FIG. 26 shows a cross-sectional view of the microtiter plate 2310along the line 25-25 of FIG. 25. The line 25-25 cuts through a row ofthe wells 2415. The wells 2415 are formed by downwardly-extending thinwalls 2510 that define the shape of the upwardly-open wells 2415. FIG.26 shows the wells 2415 having a triangular cross-sectional shape,although the wells 2415 may have other cross-sectional shapes. As isknown to those skilled in the art, a material, such as, for example, acocktail 2515 of various biological materials, can be disposed in any ofthe wells 2415 for thermal cycling. The thin walls 2510 of the wells2415 have an outer surface 2520 that contacts fluid as fluid flowsthrough the thermal cycle flow region 2345 of the flow cell assembly2315 when the microtiter plate 2310 is disposed on the flow cellassembly 2315.

[0201]FIG. 27 shows a perspective view of an exploded flow cell assembly2315, which includes an outer frame 2602 and an insert plate 2620. Theframe 2602 has an outer wall 2610 and a bottom wall 2603 that define aninterior cavity 2604 that is sized to receive the insert plate 2620. Theinsert plate 2620 includes a series of guide walls 2625 extend upwardlyfrom an upper surface the insert plate 2620. A plurality of guidebaffles 2606 extend downwardly from the insert plate 2620. The inletconduit 2325 and outlet conduit 2330 are formed by holes that arelocated in the bottom wall 2603 to provide a fluid entryway and exit wayfor the flow cell assembly, as described further below. The bottom viewof FIG. 32 shows the inlet and outlet conduit 2325, 2330.

[0202] The flow cell assembly 2315 is assembled by inserting the insertplate 2620 into the cavity 2604 of the frame 2602. FIG. 28 shows aperspective view of the assembled flow cell assembly 2315 and FIG. 29shows a top view of the assembled flow cell assembly 2315. As shown inFIGS. 28 and 29, the insert plate 2620 fits into the cavity 2604 to forman upper cavity 2615 that is sized to receive at least a portion of themicrotiter plate 2310 therein. The upper cavity 2615 defines the thermalcycle flow region 2345 (shown in FIG. 24) of the flow cell assembly2315. In the illustrated embodiment, the width of the insert plate 2620is slightly smaller than the width of the cavity 2604, so that a pair ofelongate openings are formed on either side of the insert plate 2620,one opening to form the inlet opening 2340 and the other opening to formthe outlet opening 2342. As shown in the top view of FIG. 29 andcross-sectional view of FIG. 30, the inlet opening 2340 is disposedalong a first side edge of the insert plate 2620. The correspondingoutlet opening 2342 is disposed along a second side edge of the insertplate 2620 opposite the location of the inlet opening 2340.

[0203] As shown in the cross-sectional view (along line 29-29 of FIG.28) of the is flow cell assembly 2315 in FIG. 30, the insert plate 2620forms a boundary between the thermal cycle flow region 2345 and theintermediary flow region 2335. The intermediary flow region 2335includes an inlet cavity 2805 and an outlet cavity 2810 through whichfluid can flow into and out of the flow cell assembly. The cavities2805, 2810 are peripherally surrounded by the exterior wall 2610 of theframe 2602 and enclosed on the bottom by the bottom wall 2603 of theframe 2602. As described below, fluid can flow from the inlet cavity2805 to the upper cavity 2615 of the thermal cycle flow region 2345through the inlet opening 2340, which extends through the insert plate2620. Likewise, fluid can flow into the outlet cavity 2810 from theupper cavity 2615 through the outlet passage 2342, which also extendsthrough the plate insert 2620.

[0204]FIG. 31 is a cross-sectional view of the flow cell assembly 2315along the line 30-30 of FIG. 28. As shown in FIG. 31, the inlet conduit2325 is formed by a hole in the bottom wall 2603 of the frame 2602. Theinlet conduit 2325 leads into the inlet cavity 2805 of the intermediaryflow region 2335. The outlet conduit 2330 is also formed by a hole inthe bottom wall 2603 of the frame 2602. The outlet conduit 2330 leadsinto the outlet cavity 2810.

[0205] With reference to FIGS. 28-31, the guide walls 2625 extendupwardly from the insert plate 2620 of the upper cavity 2615. The guidewalls 2625 are situated so as to form an elongate flow channel 2630between each adjacent pair of guide walls 2620. As best shown in the topview of FIG. 29 and the cross-sectional view of FIG. 30, each guide wall2625 (and corresponding flow channel 2630) is elongated and has a lengthL that extends substantially from the inlet opening 2340 to the outletopening 2342. As shown in FIG. 31, each flow channel has a height H anda width W. The height H, width W, and length L of the flow channel 2630can vary based on the microtiter plate that is used with the flow cellassembly. That is, the flow channel preferably has a width and heightsuch that the wells of the microtiter plate can fit within the flowchannel. The length L is preferably sufficiently large such that a rowof wells of the microtiter plate can be inserted into the flow channel.

[0206] As mentioned, the upper cavity 2615 is sized to receive themicrotiter plate 2310. When the microtiter plate 2310 is positionedwithin the upper cavity 2615 of the flow cell assembly, each of thewells 2415 of the microtiter plate 2310 extends downwardly into acorresponding flow channel 2630. In one embodiment, the quantity andspacing of the flow channels 2630 is substantially equal to the quantityand spacing of the rows of wells 2415 on a corresponding microtiterplate 2310. Thus, each row of wells 2415 can be inserted into acorresponding flow channel 2630 when the microtiter plate 2310 is placedwithin the upper cavity 2615 of the flow cell assembly 2315. An exampleof this is shown in FIG. 33, which shows the microtiter plate 2310positioned in the upper cavity 2615 of the flow cell assembly 2315. Whenpositioned as such, each of the six rows of wells 2415 extendsdownwardly into a corresponding flow channel 2630 of the flow cellassembly 2315. In this regard, the width W of each flow channel 2630 ispreferably large enough to accommodate insertion of a row of microtiterplate well 2315 into the flow channel 2630.

[0207]FIG. 34 shows another view of the microtiter plate 2310 positionedin the upper cavity 2615 of the flow cell assembly 2315, the view beingalong the length of one of the flow channels 2630. The microtiter plate2310 is shown in phantom lines in FIG. 34 for clarity of illustration.The length L of the guide wall 2625 that forms the flow channel 2630 ispreferably larger than the length of the corresponding row of wells 2415so that the flow channel 2630 can accommodate the entire row of wells2415.

[0208] With reference still to FIGS. 33 and 34, an upper end of theexterior frame wall 2610 can support a portion of the microtiter plate2310. A sealing ring 3110 can be positioned over the upper end of theexterior wall 2610 so that the sealing ring 3110 is interposed betweenthe upper end of the exterior wall 2610 and the microtiter plate 2310.The sealing ring 3110 can extend around the entire upper edge of theexterior wall 2610 (which surrounds the upper cavity 2615) to therebyseal the upper cavity 2615 shut when the microtiter plate 2310 ispositioned atop the side wall 2610. The sealing ring 3110 can be made ofa deformable material that conforms to shape of the upper end of theexterior wall 2610 to provide a reliable seal.

[0209]FIG. 35 is a cross-sectional view of the flow cell assembly 2315,looking downward along the line 34-34 of FIG. 30 and showing a top viewof the inlet cavity 2805 and outlet cavity 2810. A main baffle 3310forms an inlet passage 3315 of the inlet cavity 2805. The inlet passage3315 communicates with the inlet conduit 2325. As described below, afluid can flow into the inlet passage 3315 through the inlet conduit2325. The inlet passage 3315 originates at the inlet conduit 2325 (whichis located substantially in an interior of the frame 2602) and movestoward one side of the frame 2602. The inlet passage 3315 has a narrowshape and extends from the inlet conduit 2325 to a diffusion region 3317that substantially widens in size with respect to the inlet passage3315. A plurality of diffuser baffles 3320 are located in the diffusionregion 3317. The diffuser baffles 3320 are elongate and narrow in shapeand are oriented substantially parallel to the side wall 2610 of theframe 2602 so that the diffuser baffles are located at the inletopenings to the upper cavity.

[0210] With reference to FIG. 35, the main baffle 3310 also forms anoutlet passage 3324 of the outlet cavity 2810. The outlet passage 3324mirrors the shape of the inlet passage 3315. The outlet passage 3324communicates with the outlet conduit 2330 in the frame 2602. The outletpassage 3324 widens in size to form a diffusion region 3326 thatcontains a plurality of diffuser baffles 3330.

[0211] The operation of the microtiter plate assembly 2110 is nowdescribed. As discussed above, the microtiter plate assembly 2110comprises a microtiter plate 2310 that has been removably positionedatop a flow cell assembly 2315. FIG. 23 shows a plurality of microtiterplate assemblies 2110 that are positioned at thermal cycling stations2105. The operation of the microtiter plate assemblies 2110 is describedwith reference to a single microtiter plate assembly 2110, shown in FIG.36, which includes a single microtiter plate 2310 removably positionedatop the flow cell assembly 2315. The microtiter plate assembly 2110 iscoupled to an inlet pipe 2135 and an outlet pipe 2140. The inlet pipe2135 is inserted into the inlet conduit 2325 so that the inlet pipe 2135fluidly communicates with the inlet cavity 2805. The outlet pipe 2140 isinserted into the outlet conduit 2330 so that the outlet pipe 2140fluidly communicates with the outlet cavity 2810. A temperaturecontrolled fluid flows into the inlet cavity 2805 via the inlet pipe2135, as represented by the arrow 3510. The fluid originates from one ofthe reservoirs 2115 and flows to the inlet pipe 2135 via the valve andmanifold system 2130, as was described above with reference to FIG. 23.

[0212] The operation of the microtiter plate assembly 2110 is nowfurther described with reference to FIG. 37, which shows adownward-looking view of the inlet cavity 2805 and the outlet cavity2810. The temperature-controlled fluid flows into the inlet cavity 2805via the inlet conduit 2325. The fluid then flows through the inletpassage 3315 in a direction represented by the arrow 3610. The inletpassage 3315 guides the fluid toward the diffusion region 3317 of theinlet cavity 2805. The diffusion region 3317 widens in size and containsthe diffuser baffles 3320. As fluid flows through the diffusion region3317, the diffuser baffles 3320 diffuse the fluid by causing the fluidto flow through spaces between each of the diffuser baffles 3320, asrepresented by the arrows 3615. The diffuser baffles 3320 break up theflow of fluid flow and cause the fluid to evenly distribute as it flowstoward a side edge of the inlet passage 2805.

[0213] With reference now to FIG. 38, the fluid flows upwardly into theinlet opening 2340 from the inlet passage 2805, as represented by thearrow 3710. The fluid flows upwardly through the inlet opening 2340 andinto the upper cavity 2615, where the wells of the microtiter plate 2310are located. The fluid then flows through the flow channels 2630 (shownin FIG. 29) that are formed in between the guide walls 2625, asrepresented by the arrows 3720 of FIG. 38.

[0214] The fluid flow through the flow channels of the guide walls 2625is described in more detail with reference to FIG. 39, which shows a topview of the microtiter plate assembly 2110 (the microtiter plate 2310 isomitted from FIG. 39 for clarity of illustration). The guide walls 2625further diffuse the fluid flow into the separate flow channels 2630 thatare situated between each of the guide walls 2625. As represented by thebolded arrows in FIG. 39, the fluid flows in a straight line betweeneach of the guide walls 2625. Thus, the guide walls 2625 guide the fluidfrom the inlet opening 2340 toward the outlet opening 2342.

[0215] In addition to guiding the fluid from the inlet opening 2340toward the outlet opening 2342, the guide walls 2625 also guide thefluid so that it contacts the bottom surface of the wells 2415 of themicrotiter plate 2320, as shown in FIG. 38. As discussed above, thefluid is set to a predetermined temperature. The fluid can be used tocool the wells 2415 or to transfer heat to the wells 2415, depending onthe temperature differential between the fluid and the wells 2415. Inthis manner, the wells can be thermally cycled. Advantageously, theguide walls 2625 guide the fluid in such a manner that the fluid flowsin a straight line over the wells 2415, thereby eliminating uneven orturbulent fluid flow over the wells of the microtiter plate. Thisprovides for a more even heat transfer between the fluid and themicrotiter plate. The guide walls 2625 also ensure that fluid contactsall of the wells of the microtiter plate.

[0216] With reference still to FIG. 38, the fluid next flows downwardlyinto the outlet opening 2342 from the upper cavity 2615, as representedby the arrow labeled 3725. The fluid flows downwardly through the outletopening 2342 into the outlet cavity 2810, as represented by the arrowlabeled 3720. The fluid flow through the outlet cavity 2810 is nowdescribed with reference to FIG. 37. Once the fluid enters the outletcavity 2810, the fluid flows around the diffuser baffles 3330 toward theoutlet passage 3324, as represented by the arrows labeled 3620. Thefluid enters the outlet passage 3324 and flows into the outlet conduit2330, as represented by the arrow labeled 3630.

[0217] With reference now to FIG. 36, the fluid exits the outlet cavitythrough the outlet conduit 2140, as represented by the arrow labeled3520. As discussed above, the outlet conduit 2330 fluidly communicateswith the outlet pipe 2140 (shown in FIGS. 23 and 36). The fluid flowsinto the outlet pipe 2140, which guides the fluid back into theappropriate reservoir 2115 via the valve and manifold system 2130, shownin FIG. 23.

[0218] Plate Sealing

[0219] The microtiter plates used with the automated process line 100are typically sealed with an aluminum or polypropylene adhesive film.This prevents evaporation during thermal reactions. But it is possibleto get some condensation of solution on the inside of the seal.Therefore, the plates are subjected to a centrifuge so that the solutioncollects at the bottom of the microtiter plate wells, although there isstill a very small chance of some sample collecting on the inside of theseal. When the seal is removed, it is important that there be no crosscontamination of samples. To avoid this, the system 100 uses a “peeler”comprising a robotic arm. The seal for the microtiter plates is designedto be bigger than the plate, and a portion of the sealing film extendsout from the plate on the short axis (or it may be on the long axis if adifferent movement of the robotic arm is configured). The microtiterplate, while moving down the conveyor, is stopped at a defined positionand there the plate is the gripped and held steady.

[0220] A robotic arm with a different gripper that has fingers which cantouch each other then maneuvers, such that the gripper fingers locatethe film and tighten, and so grip the film. The robotic arm then raisesslightly and then moves along the length of the microtiter plate. As itmoves it pulls the sealing film with sufficient force so as to break theadhesive pull that the film has for the microtiter plate. The grippermoves at such a height as to ensure that the originally inward side ofthe seal is now pointing upward away from the remainder of the sealedmicrotiter plate. The angle of the removing seal is such that should anydroplets of solution or sample be on the inside of the seal that it doesnot move down the surface of the film and therefore possibly back into adifferent open well of the microtiter plate. The nature of the filmsurface is chosen to have sufficient surface tension for the solution orsample being used to ensure minimal or ideally no movement of a dropleton the film except at an extreme angle or force not typicallyencountered.

[0221] The present invention has been described above in terms of apresently preferred embodiment so that an understanding of the presentinvention can be conveyed. There are, however, many configurations forsample handling systems not specifically described herein but with whichthe present invention is applicable. The present invention shouldtherefore not be seen as limited to the particular embodiments describedherein, but rather, it should be understood that the present inventionhas wide applicability with respect to sample handling generally. Allmodifications, variations, or equivalent arrangements andimplementations that are within the scope of the attached claims shouldtherefore be considered within the scope of the invention.

What is claimed is:
 1. A particle dispensing device having a proximalend and a distal end, said particle dispensing device comprising: ahollow tube having a proximal end with an opening, a distal end with anopening, and an inside surface; a plunger slideably inserted through theproximal end of the hollow tube; and one or more particles of solidreagent disposed within the distal end of the hollow tube.
 2. Theparticle dispensing device of claim 1, wherein the proximal end of theparticle dispensing device is configured to be coupled to a dispensingassembly comprising an array of particle dispensing devices.
 3. Theparticle dispensing device of claim 1, wherein the plunger comprises aproximal end and a distal end, and wherein a space is formed between thedistal end of the plunger and the distal end of the hollow tube, saidspace defining a predetermined volume sufficient for containing said oneor more particles of solid reagent.
 4. The particle dispensing device ofclaim 1, wherein the plunger has a proximal end and a distal end, saidproximal end configured to be coupled to a flat plate, and said distalend in contact with said one or more particles of solid reagent.
 5. Theparticle dispensing device of claim 1, wherein at least one of the oneor more particles of solid reagent are frictionally engaged with theinside surface of the hollow tube.
 6. The particle dispensing device ofclaim 1, wherein the hollow tube and plunger are both cylindrical. 7.The particle dispensing device of claim 1, wherein the plunger is longerthan the hollow tube.
 8. The particle dispensing device of claim 1,wherein the solid reagent comprises resin.
 9. The particle dispensingdevice of claim 1, wherein a tight seal is formed between an outsidewall of the plunger and the inside surface of the hollow tube.
 10. Theparticle dispensing device of claim 9, wherein the plunger comprises adistal end with a channel formed on its outer surface and an o-ringmounted coaxially in the channel.
 11. A particle dispensing devicehaving a proximal end and a distal end, said particle dispensing devicecomprising: a hollow tube having a proximal end with an opening, adistal end with an opening, and an inside surface; and a plungerslideably inserted through the proximal end of the hollow tube, whereinthe proximal end of the particle dispensing device is configured to becoupled to a dispensing assembly comprising an array of particledispensing devices.
 12. The particle dispensing device of claim 11,wherein the plunger comprises a proximal end and a distal end, andwherein a space is formed between the distal end of the plunger and thedistal end of the hollow tube, said space defining a predeterminedvolume sufficient for containing one or more particles of a solidreagent.
 13. The particle dispensing device of claim 11, wherein theplunger has a proximal end and a distal end, said proximal endconfigured to be coupled to a flat plate.
 14. The particle dispensingdevice of claim 11, wherein the hollow tube and plunger are bothcylindrical.
 15. The particle dispensing device of claim 11, wherein theplunger is longer than the hollow tube.
 16. The particle dispensingdevice of claim 11, wherein said solid reagent comprises resin.
 17. Aparticle dispensing device having a proximal end and a distal end, saidparticle dispensing device comprising: a cylindrically shaped hollowtube having a proximal end with an opening, a distal end with anopening, and an inside surface; a cylindrically shaped plunger slideablyinserted through the proximal end of the hollow tube, said plungerhaving a proximal end and a distal end, the proximal end of the plungerconfigured for coupling to a flat plate, wherein a space is formedbetween the distal end of the plunger and the distal end of the hollowtube, said space defining a predetermined volume; and one or moreparticles of a solid reagent substantially filling the space formedbetween the distal end of the plunger and the distal end of the hollowtube, wherein the proximal end of the particle dispensing device isconfigured to be coupled to a dispensing assembly comprising an array ofparticle dispensing devices.
 18. The particle dispensing device of claim17, wherein the solid reagent comprises resin
 19. The particledispensing device of claim 17, wherein the distal end of the plungercomprises a channel formed on its outer surface and an o-ring mountedcoaxially in the channel.
 20. A method of dispensing a solid reagentcomprising: providing a particle dispensing device comprising: a hollowtube having a proximal end with an opening, a distal end with anopening, and an inside surface; and a plunger having a proximal end anda distal end, said plunger slideably inserted through the proximal endof the hollow tube; positioning the plunger within the hollow tube sothat a space is formed between the distal end of the plunger and thedistal end of the hollow tube, said space defining a predeterminedvolume; positioning the distal end of the hollow tube over a reservoircomprising one or more particles of said solid reagent; forcing thedistal end of the hollow tube into the reservoir, thereby filling thespace with one or more particles of solid reagent; positioning theparticle dispensing device over a solid support; and using the plungerto push the one or more particles of solid reagent out into the solidsupport.
 21. A method as defined in claim 20, wherein the one or moreparticles of solid reagent are frictionally engaged with the insidesurface of the hollow tube after the hollow tube is lowered into thereservoir.
 22. A method as defined in claim 20, wherein the proximal endof the plunger is coupled to a flat plate, the flat plate being apredetermined distance from the hollow tube.
 23. A method as defined inclaim 22, wherein pushing the one or more particles of solid reagent outinto the solid support is performed by moving the flat plate toward thehollow tube.
 24. A method as defined in claim 22, wherein pushing theone or more particles of solid reagent out into the solid support isperformed by moving the hollow tube toward the flat plate.
 25. A methodas defined in claim 20, further comprising packing the solid reagentwithin the reservoir prior to the step of lowering the hollow tube intothe reservoir.
 26. A method as defined in claim 20, wherein forcing thehollow tube into the reservoir is performed by lowering the hollow tubeinto the reservoir.
 27. A method as defined in claim 20, wherein forcingthe hollow tube into the reservoir is performed by raising the reservoirtoward the hollow tube until the hollow tube is forced into thereservoir.
 28. A method as defined in claim 20, wherein the solidreagent comprises resin.
 29. A method as defined in claim 20, whereinthe solid support comprises a microtiter plate.